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THE
ACTINOMYCETES
VOLUME TI
Ferdinand Cohn (1828-1898), who was the first to observe and describe an actino-
mycete (1875), under the name Streptothriz Foerstert.
THE ACTINOMYCETES
Volt
CUA ssh CATLON 2 DENY EEDCA TON AN D
DESGHRIPTIONS OF GCENERA AND SPECIES
by
Selman A. Waksman
BALTIMORE
iE Web ieh FAV Sivek WEE KILNS GCOMPAN Y
1961
THE ACTINOMYCETES
Vou. II: CLASSIFICATION, IDENTIFICATION AND
DESCRIPTIONS OF GENERA AND SPECIES
Copyright ©, 1961
The Williams & Wilkins Company
Made in the United States of America
Library of Congress
Catalog Card Number
59-9962
Composed and printed at the
WAVERLY PRESS, INC.
Baltimore 2, Md., U.S.A.
PREFACE
In 1922, Professor D. H. Bergey of the University of Pennsylvania wrote to
me that he and the Committee on Characterization and Classification of the
Society of American Bacteriologists were in the process of preparing a ‘‘Manual
of Determinative Bacteriology’’; he asked whether I would be willing to under-
take the preparation for that volume of a section dealing with the actinomycetes.
This group of organisms had occupied my attention for the previous several
years, and alone (1919) and with Roland Curtis (1916), I had deseribed a num-
ber of new species; yet I hesitated to accept this assignment. There were several
important reasons for this hesitation: (a) I was not at all sure that the descrip-
tions of actinomycetes so far published provided sufficient information for the
accurate identification of most of the species recorded in the literature; (b) only
four years previously, | had been warned by the dean of American cryptogamic
botanists, Roland Thaxter, not to make further descriptions of new species based
solely or largely upon cultural and biochemical properties; and finally (¢) I was
not even certain at that time whether the actinomycetes should be included
with the bacteria.
I told Professor Bergey all this and suggested that it would be better to wait
a few years until more detailed information was obtained concerning this group
of microorganisms, especially with regard to their morphological and biochemical
properties, before an attempt was made to codify them. I received a curt and
somewhat sarcastic reply that if I would not, for one reason or another, under-
take this task, he would have to do it himself. My immediate answer was, ‘‘I
will do it.”? The best that I could accomplish at that time was to use cultural and
biochemical characteristics as a major basis for the classification of the actinomy-
cetes and for the characterization of the known species.
Since then, or for more than a third of a century and for seven consecutive edi-
tions of ““Bergey’s Manual,” I have been largely responsible for the preparation
of the descriptions of the actinomycetes. I have not, however, always had the
final word in organization of the material for all the various editions. Alone
(1940), and together with Professor A. T. Henrici of the University of Minnesota
(1943), I proposed two systems for classification of the actinomycetes, the second
of which consisted of a thorough revision of the group and its separation into four
genera. The most significant change in this revision was the proposal, in 1943, of
the new generic name, Streptomyces. This second system has been the basis for
the organization of the material in the last two editions of Bergey’s Manual.
In presenting this volume, I am now certain of one thing, namely, that the
place of the actinomycetes is definitely among the bacteria and not among the
fungi. Ample evidence of this belief has been presented in Volume I of this trea-
tise. Unfortunately, the first reason for my hesitancy in 1922, I believe, remains
valid; the accuracy of the information available for species identification is still
Ve
vl THE ACTINOMYCETES, Vol. II
open to question. The chief reason for this uncertainty is that although much
knowledge has since accumulated, especially during the last 20 years when many
Streptomyces species became known as antibiotic-producing organisms, taxonomic
work was largely neglected except by a few dedicated investigators. Recently,
however, several important contributions (Hesseltine et al., 1954; Flaig and
Kutzner, 1954; Kutzner, 1956; Waksman, 1957; Ettlinger et al., 1958; Pridham,
1959) to this subject have appeared. A survey of the recent literature shows that
morphological characters are tending to replace physiological and cultural proper-
ties as the leading criteria in species characterization. It may be said that we are
now in a transitional stage in which our ideas are changing, not only concerning
the usefulness of criteria for species differentiation, but also with regard to the
species concept. Since a classification of a group of living organisms is always only
‘preliminary,’ based upon the current knowledge of these organisms, I believe
that, in summarizing the subject at present, and in trying to combine the older
and newer ideas, I have presented useful criteria for species differentiation and
an outline of species concept for the genera Actinomyces, Nocardia, Streptomyces,
Micromonospora, and certain others.
The rapidly accumulating information about the separation of some of the
genera into distinct groups or sections, the recent introduction of several new
genera, and the description of numerous new species, all necessitated a complete
recasting of the material presented in the last edition of Bergey’s Manual and in
other treatises. This volume is largely the result. An attempt has been made to
bring together in this volume all the information required for the identification
of newly isolated cultures of actinomycetes. All descriptions and names for which
insufficient data have been provided, especially when no reproducible media have
been employed, have been placed in a separate chapter as ‘‘incompletely de-
scribed.”? Descriptions in which excessive and often confusing information has
been presented, have been abbreviated to fit a certain ‘‘standard.”’ Often, this
standard has turned out to be a Procrustean bed. I beg forgiveness, both from
the ‘‘reader” and from the preservers of the Code (International Code of Nomen-
clature of Bacteria and Viruses). My sole apology is that it is my sincere hope
that it would serve the purpose.
The author wishes to acknowledge his sincere indebtedness to Dr. Norvel M.
McClung of the University of Georgia, to Dr. R. E. Buchanan of Iowa State
University, and to Dr. Ruth E. Gordon and Dr. Hubert A. Lechevalher of this
Institute, for reading individual chapters and for making valuable suggestions;
to Dr. Hans J. Kutzner of this Institute and Dr. Thomas G. Pridham of the
Northern Regional Research Laboratory, for reading the major portions of this
volume and for suggesting numerous corrections and modifications; to Miss Alma
Dietz of the Upjohn Company, Dr. Edward J. Backus of the Lederle Labora-
tories, and all others who kindly supplied photographs; to Mrs. Herminie B.
Kitchen for editorial work, and to Mr. Robert A. Day for assistance in the prep-
aration of the various illustrations and for reading the entire manuscript,
Selman A. Waksman
INTRODUCTORY
This volume deals exclusively with the well recognized genera of the actino-
mycetes. No consideration is given here to the various closely related genera that
are often included in the order Actinomycetales, notably the genus Mycobacterium
Lehmann and Neumann, 1896.
The actinomycetes comprise three families, which are further subdivided into
LO genera.
A. Spores formed, but not in sporangia.
I. Vegetative mycelium fragmenting into bacillary or coccoid elements.
Family I. Actinomycetaceae Buchanan.
1. Anaerobic or microaerophilic, nonacid-fast.
Ll. Actinomyces Harz
2. Aerobic, partially acid-fast or nonacid-fast.
2. Nocardia Trevisan
II. Vegetative mycelium nonseptate, not fragmenting into bacillary or coc-
coid elements.
Family II. Streptomycetaceae Waksman and Henrici.
1. Aerial mycelium produced.
a. Spores formed in chains.
3. Streptomyces Waksman and Henrici
b. Spores formed singly.
4. Thermoactinomyces Tsiklinsky
c. Spores occurring in pairs or in chains.
a'. Mesophilic forms, in pairs.
5. Waksmania Lechevalier and Lechevalier
b!. Thermophilic forms, in pairs or in chains.
6. Thermopolyspora Henssen
2. Aerial mycelium not produced.
a. Spores occurring singly on short sporophores.
al. Mesophilic forms.
7. Micromonospora Orskov
b!. Thermophilic forms.
8. Thermomonospora Henssen
B. Spores occurring in sporangia.
Family III. Actinoplanaceae Couch
I. Aerial mycelium usually not formed, coiled conidial chains lacking, spor-
anglospores motile.
9. Actinoplanes Couch
II. Aerial mycelium abundant, coiled conidial chains as well as sporangia
formed in some species, sporangiospores nonmotile.
10. Streptosporangium Couch
vil
vill THE ACTINOMYCETES, Vol. IT
Certain other genera, recently suggested, have been given tentative considera-
tion.
These genera comprise about 350 species. In addition to these, a large number
of other species are listed as ‘incompletely described.”
TABLE OF CONTENTS
The Actinomycetes
VoLuME II
CLASSIFICATION, IDENTIFICATION, AND DESCRIPTION
OF GENERA AND SPECIES
PTE RC Cah Oreo Se Ba wig es. RPSL Aci: Ste Pant ess A Bn sk a a I Ae tne V
MNT OCCU OTS chance co o.8 Soi a Pe te, a wna Neer Gik-oth a MeL eee vil
1. The Species Concept in Relation to the Actinomycetes.............. |
Deal CUE E MUSA CLEMOMUCES 2:0 See or Pale a heaas ves seas Foe oes eee tad ee exe 12
“ah IU OEM (ey NILOTSENA'A(O170 1410 110 NEA ee Pe oe Be a oes ot mE cm ne 21
4. Characterization of Streptomyces Species.............. 02.0000 eee 61
5. Systems of Classification and Identification of Groups and Species of the
GSMS HSU DLONUNCCG tice ole gts oie lois ijt Fe Boas e SeOPO ta Ae ne oe oR 82
6. Series and Species of the Genus Streptomyces... ........0.0...0.000-. LS
fe Classiication ob wieplomiyces SPeCles.. - 2.6 ce. bebe ecw ove see nies 152
S. Deseriptionror Species of Streptomyces... o.oo wee ne 165
Omiihe: Genus Mecromonospordy, les ele. Slee els ca da See lasagne saan 293
LOMeithesGenusaw aksmanica (Microbtspora)): co ..08 2 fee le hs oso ed Ss 298
ea hiermopnilicy Actinomycetes. ..tsS sob. ae Oe bs bo awlegan eee ba bs 300
WAAC HINO DIAM ACCACUe +. oes teas es es Be aoe 2 eee 310
13. Incompletely Described Species of Actinomycetes................... 315
Appendix I. Color Designations for Describing Actinomycetes (Lin-
GLEMDEII) Eee Ee UPA AI Mente sae. Slee CEs EO OS oat te eee 327
Appendix II. Certain Important Media for the Study of Actinomycetes 328
| 572) (EN 2) CET ARN Oa mane ae Maen Se rd OS eee ae ee) AE 335
InICexXAG MOTE AIMISMIS SSeS eo ENA Ee cl Meee 347
Crenerall Sinden... Mle tice, at teas: syns Neen e cae Uy cake mute clog CR Lt een eee 360
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Chapter |!
The Species Concept in Relation to
the Actinomycetes
Systematic Position of the Actinomy-
cetes
In the preface to this volume, the state-
ment was made that ‘I am now certain of
one thing, namely, that the place of the
actinomycetes is definitely among the bac-
teria and not among the fungi. Ample evi-
dence of this belief has been presented in
Volume I of this Nevertheless,
some reiteration is warranted at this point.
The taxonomic position of the
mycetes, notably their relationship to the
bacteria, on the one hand, and to the fungi,
on the other, has been one of the most de-
batable questions in microbiology. The size
(width of thallus) and staining properties of
the actinomycetes have usually placed them
with the bacteria. Their branching and man-
ner of sporulation have suggested their rela-
tionship to the fungi. Still other properties of
treatise.”’
actino-
actinomycetes seemed to warrant their con-
sideration as a transition group between the
bacteria and the fungi.
tecent evidence seems to point definitely
to the fact that the actinomycetes are more
closely related to the bacteria:
1. Some of the actinomycetes, such as
species of Actinomyces and Nocardia, are
closely related to true bacteria, notably spe-
cies of Lactobacillus and Corynebacterium.
2. Neither
have been shown to contain true nuclei; they
both contain only chromatin granules dis-
tributed through the hyphae or the cells.
actinomycetes nor bacteria
3. The diameter of actinomycete my-
celium and spores is similar to that of bac-
teria. Actinomycetes also, as a rule, lack
septa.
4. Actinomycetes are subject to attack by
phages just as bacteria are; filamentous
fungi are not.
5. Actinomycetes are usually sensitive
(allowing for strain variability) to antibiotics
that are active upon bacteria; they are usu-
ally resistant to those antibiotics, like the
polyenes, that are active upon fungi but not
upon bacteria.
6. Chitin is absent from the cell substance
of actinomycetes as well as from bacterial
cells, but is present in fungus mycelium and
In their lack of cellulose, actino-
mycetes are also similar to most bacteria and
unlike fungi. Avery and Blank (1954) con-
cluded that “from the chemical point of view
Actinomycetales have nothing in common
with the true fungi, but rather with the bac-
Cummins and Harris (1958)
even further by suggesting that the order
spores.
teria.”’ went
Actinomycetales be abolished altogether and
that the families of the actinomycetes be
included in the Hubacteriales.
7. Like bacteria, but unlike most fungi,
actinomycetes as a rule are sensitive to
an acid reaction of the medium.
8. The close relationship of the actino-
mycetes to the bacteria is also evident from
the work of Couch (1954), who found that
certain J/icromonospora-like forms resemble
2 THE ACTINOMYCETES, Vol. II
those of bacteria. Couch emphasized the re-
semblance of the mycelium and sporangia of
Actinoplanes to those of the chytrids; he con-
cluded that this genus may represent a con-
necting link between the bacteria and the
lower fungi.
The Generic Problem with Actinomy-
cetes
Prior to 1943, several systems of classifi-
cation of actinomycetes had been proposed.
In most instances, all the species were in-
cluded in a single genus, which was fre-
quently designated by different names. The
most common of these names were the two
oldest, Streptothrix and Actinomyces. <Al-
though occasional efforts had been made to
separate the actinomycetes into several gen-
era, such attempts usually failed to receive
more than passing attention. The work of
Waksman (1919), @rskov (1923), Jensen
(1931), and Erikson (1935) finally led Waks-
manand Henrici to suggest, in 1948, the divi-
sion of the actinomycetes into four genera. A
new genus, Streptomyces, was proposed to in-
clude those forms that are characterized by
the production of an aerial mycelium with
catenulate spores. Most of the important an-
tibiotic-producing organisms subsequently
have been found to belong to this genus.
Unfortunately, this generic separation
brought with it a number of new problems,
which can be briefly summarized as follows:
1. There is considerable overlapping
among the different genera, notably between
certain forms of Streptomyces that have lost
the capacity to produce aerial mycelium and
species of Nocardia, as brought out in a re-
cent paper by Gordon and Smith (1955);
there is also overlapping between certain
nocardiae and mycobacteria.
2. The formation by species of Strepto-
myces and by certain forms of Nocardia of
two different types of mycelium, substrate
and aerial, and the influence of previous con-
ditions of cultivation upon the growth and
biochemical activities of these organisms
served to confound the existing confusion.
The nomenclatural status of the genera of
Actinomycetales has recently been discussed
by Lessel (1960).
Lechevalier et al. (1961) described a new
genus Micropolyspora (type species M. bre-
vicatena), an organism that fragments like
the members of the family A ctinomycetaceae
and sporulates like a member of the Strepto-
mycetaceae, by forming chains of conidia on
aerial hyphae; it also forms chains of conidia
on the substrate mycelium. These authors
suggested that the family Streptomycetaceae
be dropped and the family A cténomycetaceae
be enlarged to include the genera Actinomy-
ces, Muicromonospora, Thermoactinomyces,
Waksmania, Micropolyspora, Nocardia, and
Streptomyces.
What Is a Microbial Species?
In the study of the taxonomy of any group
of living organisms, including microorgan-
isms, one is faced sooner or later with the
problem of defining what is meant by a
species. With microorganisms, in usual prac-
tice, a microbial culture is designated by
a name, sometimes qualified with a strain
number; its morphological and cultural
properties, and frequently its ecological and
etiological characteristics, are described suf-
ficiently so that anyone who finds this or-
ganism in nature will be able to recognize it
from the description. If possible, the type
form of the species is preserved in a type
culture collection, to aid in the future identi-
fication of the species.
Unfortunately, microbial forms and types
of organisms are not fixed in nature or even
in culture. Some strains, even those closely
related to the fixed type, may differ enough
to raise a question as to their exact or specific
identity. This frequently leads, often on the
basis of only minor differences, to the crea-
tion of new species that are given new epi-
thets. This is particularly true of those mic-
SPECIES CONCEPT IN RELATION TO ACTINOMYCETES
roorganisms, like the actinomycetes, that
occur abundantly in nature; some of the
newly isolated cultures may differ greatly
from the fixed types. The difficulty of estab-
lishing and recognizing ‘“‘species’”’ under these
conditions may become particularly perplex-
ing. Raper (1954) was fully justified in say-
ing, “It is almost axiomatic that the ease
with which a species of microorganism can be
recognized tends to vary inversely with the
number of isolates available for observation
and examination.”
The concept of ‘“‘species”’ first used during
the seventeenth century gradually came to
denote the fundamental units of a biological
classification. These units came to be re-
garded as fixed or static entities, created by
nature, which can be grouped into higher
categories, namely, genera, orders, and
classes. As the evolutionary theory was grad-
ually accepted, especially with the develop-
ment of modern genetics and cytology, the
concept of ‘‘species” began to undergo a
change.
Hucker and Pederson (1931) emphasized
that the difficulty of dividing lower forms
into well-defined species has led many to
question whether these are natural groups
and whether they can be considered to be
similar to ‘“‘species’”? among higher forms of
life. The problem always arises: How much
difference must exist between two cultures
of bacteria before we are justified in regard-
ing them as distinct species?
Krassilnikov (1938) was very emphatic in
stating that many investigators, without
considering the rules of nomenclature pro-
posed at international congresses, either de-
scribe the same forms under different names
or combine various organisms into the same
species. He said: ‘Even the concept of
‘species’ is considered differently by various
workers depending on their individual point
of view, frequently considering a minor lack
of correlation of a certain character as suffi-
cient justification for creating a new species.”
oe
ae
Just as in the case of many groups of true
bacteria, one of the causes of the chaotic
state of nomenclature of the actinomycetes
is the lack of type cultures. It has actually
been suggested (Skerman, 1949) that even
the available cultures be completely rede-
scribed, priorities being based on existing
names, and those names and descriptions for
which no type cultures are available be dis-
sarded.
In comparing the species concept among
microbes with that of higher plants and
animals, Cowan (1956) suggested that con-
sideration be given to the following aspects:
(a) whereas larger plants and animals have
geographical distribution areas, few microbes
have such particular areas; (b) morphology
is essential for the separation of species
among algae, fungi, and protozoa, but. it
barely distinguishes higher ranks among bac-
teria; (c) cytology is useful at the generic
level, but ‘‘at the species level the bacteriol-
ogist relies more on physiological than on
morphological differences”; (d) interfertility
is hardly to be considered as a species char-
acter, since bacteria and actinomycetes re-
produce asexually; (e) the introduction of
certain characters in microbiology not uti-
lized by botanists and zoologists adds satis-
factory classification criteria; these include
“nutritional requirements, metabolic and
‘atabolic products, antigenic structure and
pathogenicity.”
In discussing bacterial classification,
Sneath (1957) came to the following con-
clusions: (a) an ideal classification is one
which has the greatest content of informa-
tion; (b) over-all similarity is the basic con-
cept of such an ideal classification, and is
measured in terms of the number of similar
features possessed by two organisms; (c)
every feature should have equal weight; (d)
the division into taxonomic groups is made
upon correlated features.
To avoid the growing confusion from con-
flicting ideas, Gilmour (1958) suggested sep-
4 THE ACTINOMYCETES, Vol. II
aration of the concepts of “nomenclatural
taxonomy” from those of ‘experimental tax-
onomy.”’ It is to be remembered that species
are, after all, convenient ‘“‘artificial creations
of human imagination” rather than “‘real
biological entities.”” Gilmour further sug-
gested that “nomenclatural categories of
genus, species, variety, etc.” are excellently
suited for the purpose of ‘‘a broad map of
the diversity of living things.” It would,
therefore, be ‘‘a great advantage if they were
not subject to continued attempts to bring
them up to date and to redefine them in
evolutionary terms.”
Speciation of Actinomycetes Other than
Streptomycetes
Kxrassilnikov (1938) wrote, ‘In spite of the
most extensive literature, we have no definite
idea concerning the natural systematics of
the actinomycetes, nor a single opinion of
their structure and development.’ The re-
cently accumulated information leads us to
conclude, however, that we need not be so
pessimistic.
According to Pridham (1959), there are
now known more than 100 genera of actino-
mycetes and well over 1500 subgeneric names
and specific, or subspecific, epithets. Some of
the descriptions of these forms are good,
others lack essential details, and many are
worthless. Morphological criteria are be-
lieved to play an important role in separa-
tion at the generic level (Fig. 1), with a
gradual intergradation in complexity of re-
productive units. The actinomycetes are
looked upon as a heterogeneous group of
organisms, ranging from the simple myco-
coeci and the seemingly more complex no-
vardiae to the straight or flexuous strepto-
mycetes and the verticillate forms, and from
the relatively simple micromonosporae to
forms such as Waksmania, Actinoplanes, and
Streptosporangium (the latter two genera pos-
sibly having some affinities with the chy-
trids). Some of these organisms have definite
affinities with true bacteria, others with both
bacteria and microfungi, and still others with
phycomycetous fungi.
This heterogeneity is further emphasized
by the facts that the actinomycetes contain
forms that are anaerobic, microaerophilic, or
aerobic; forms that fragment and those that
do not; and forms that produce aerial my-
celium and those that do not. Pridham sug-
gested that some of the present concepts
centered around the three genera Actino-
myces, Nocardia, and Streptomyces be ac-
cepted. Thus included in the Actinomyces
would be the anaerobic to microaerophilic
forms; in the Nocardia, the aerobic types
that either form no aerial mycelium or pro-
duce an aerial mycelium that generally has
no catenulate spores; and in the Streptomy-
ces, the aerobic forms that generally produce
satenulate spores.
Although time and again taxonomists have
emphasized that an effective system of clas-
sification should be based upon criteria that
are expressed in consistently reproducible re-
sults, this has hardly been applied, at least
so far as our present knowledge is concerned,
to the species characterization of actino-
mycetes. Many ‘‘new species’”’ have been de-
seribed on the basis of a single difference—
frequently a quantitative variable—from
“old species.”? One often wonders what the
composition of the medium, the conditions of
growth, and the natural variability observed
so frequently among duplicate cultures have
to do with these distinguishing properties.
The species concept among the actinomy-
cetes must be considered as the continuity
between different groups of organisms desig-
nated as species, with various transitional
forms bridging the gaps between species. The
concept of natural classification apples to
actinomycetes perhaps better than to many
other bacterial groups: there are the chemi-
cal approach (chemical composition, pres-
ence of specific chemical compounds), the
morphological approach (type of aerial my-
SPECIES CONCEPT IN RELATION TO ACTINOMYCETES 5)
Mycococcus 7
Jensenia l \7
Polysepta ye Lee
Mycobacterium ('\ Y
Actinomyces } fe
A
; é
Nocardia
Proactinomyces
Pseudonocardia
B
Streptomyces
Chainia®
Streptoverticillium*
Micromonospora
Thermoactinomyces
Thermomonospora ee
Microbispora?
Thermopolyspora;
Waoaksmania
ee
Actinoplanes yz - wo
Streptosporangium Af
Fraure 1. Morphology of the various genera of the Actinomycetales. Of these, only Actinomyces in
A, Nocardia in B, Streptomyces in C, Micromonospora, Thermoactinomyces, Waksmania. Actinoplanes
i 4 I q I )
and Streptosporangium in D are recognized in this treatise as true actinomycetes; Nocardia and Proac-
tinomyces are synonyms (Courtesy of T.
Research Service, U. S. Department of Agriculture).
G. Pridham of the Northern Regional Laboratory, Agriculture
6 THE ACTINOMYCETES, Vol. II
celium, type of sporulation, shape and sur-
face of spore), and finally the ecological ap-
proach (anaerobie versus aerobic, pathogenic
versus nonpathogenic, thermophilic versus
mesophilic). The idea of a physiological clas-
sification includes formation of antibiotics
and of enzymes, utilization of carbon com-
pounds, and transformation of nitrogenous
compounds, all of which can supply supple-
mentary information.
Speciation of Streptomycetes
What has been said for the actinomycetes
as a whole applies particularly to the large,
heterogeneous, and variable group of organ-
isms represented in nature by the aerial my-
celium-producing strains, most of which are
included at present in the genus Streptomy-
ces. These organisms are found in the soil in
the form of hundreds of thousands of spores
and of bits of mycelium per gram. They are
also found extensively in manures and in
composts, in various fresh-water basins, in
dust, and on food. They are almost entirely
absent from peat bogs and the sea.
The actinomycetes belonging to the genus
Streptomyces have recently come to occupy
an eminent place because many of them are
important producers of antibiotics, vitamins,
and enzymes.
With the growing economic significance of
members of the genus, the establishment for
each species of certain characteristics which
would be adequate to enable the investigator
to recognize freshly isolated cultures in well
defined specific terms becomes of great theo-
retical and practical importance.
Following the first descriptions of Cohn
(1875), very few additional species of the
aerial mycelium-producing actinomycetes
were recognized until 1914. This was true in
spite of the rapidly accumulating literature
on the occurrence of such actinomycetes in
the soil and in the causation of plant diseases.
The designations limited
largely to the names ‘‘Actinomyces albus”’
common were
and ‘‘Actinomyces chromogenus,” depending
on the color of the aerial mycelium or the
formation of soluble, dark pigments in com-
plex organic media.
Rossi-Doria (1891) was the first to de-
scribe an organism, under the name Strepto-
thrix alba, which was later designated as the
type of the genus Streptomyces proposed by
Waksman and Henrici in 1943. The most
important characteristics of this species are
its white aerial mycelium and the tendency
for colonies to form concentric rings of this
aerial mycelium. Rossi-Doria noted the abil-
ity of his organisms to grow on numerous
complex organic substrates.
Thaxter (1891), who first described an im-
portant economic species, the causative
agent of potato scab (which he believed to
be a fungus, Oospora), was highly critical of
the efforts to describe ‘‘species”’ largely on
the basis of cultural properties of the organ-
isms. In this respect, the actinomycetes do
not differ from any of the other groups of
bacteria, where cultural properties and bio-
chemical reactions have to supplement in-
sufficient morphological information. Physi-
ological activities and ecological properties,
which are the expression of the response of
organisms to their environment, are too
numerous and often too variable among ac-
tinomycetes to justify unlimited confidence.
Krainsky (1914), Waksman and Curtis
(1916), and Waksman (1919) emphasized
the use of synthetic substrates, mn addition
to organic media. Carbon and nitrogen utili-
zation tests were employed. Added attention
was given to micromorphology. Many new
species were described. Jensen (1930a, 1931)
and Duché (1934) added various new species,
the latter investigator stressing the use of
various combinations of carbohydrates and
nitrogenous compounds as media ingredients.
One of the reasons for the limited recog-
nition of species among the aerial mycelium-
producing actinomycetes prior to 1914 was
the fact that protein-rich media were
SPECIES CONCEPT IN RELATION TO ACTINOMYCETES
employed for their cultivation. With the in-
troduction of synthetic media, it became
definitely established that the aerial myce-
lium-producing actinomycetes comprise a
large number of forms, differing greatly in
their physiological and biochemical proper-
ties, and to a lesser degree in their morphol-
ogy. It was also recognized that, if a suffi-
ciently large number of cultures was isolated
and examined, many differences would be
noted suggesting variability of the type spe-
cies. The concept ‘‘species-groups,”’ with one
culture as the type species, was suggested.
Waksman (1919) emphasized, therefore, that
in spite of variation of individual biochemi-
cal characteristics of the actinomycetes,
there are certain well defined properties, no-
tably morphology, color of aerial mycelium,
and formation of soluble pigments, that char-
acterize these organisms, especially when
grown on standard synthetic media and un-
der carefully controlled conditions of tem-
perature and aeration.
It is easy to pick out a few cultures of
actinomycetes (or streptomycetes) which
possess characteristic properties that can be
recognized as distinct species, and to discard
all the others. This was actually done by
Waksman and Curtis in their early (1915-
1916) classification of actinomycetes, since
they were faced with such a large number of
freshly isolated cultures that it was impos-
sible to consider more than a very small
number of them. How many others have
acted likewise it is difficult to say. Should
the various intermediate strains be consid-
ered, one might be inclined to regard each
as a different species, distinct from the
others in at least one variable property, be
it morphological, cultural, or biochemical.
With the examination, in recent years, of
many thousands of cultures of actinomy-
cetes for their antibiotic properties, such an
attitude was frequently reduced to an ab-
surdity. There are those who contend that
the insistence on permanent characteristics,
~I
preferably a group of them, in describing new
species, would limit greatly our recognition
of the growing economic importance of these
organisms. Then there are those who reason
that not enough species of actinomycetes
have so far been described, thus justifying
random descriptions of many freshly isolated
strains as new species.
Even synthetic media did not yield the
final answer to the species problem of this
group of organisms. Their cultural proper-
ties, or growth characteristics in media of
different chemical composition, properties
that were at first greatly emphasized, were
found to be extremely variable. Type cultures
were shown to change their specific charac-
teristics when grown in artificial media.
Saltations and mutations came to play a
highly important part in changing such prop-
erties. When morphology was recognized at
all, it was limited largely to observations on
the curvature of the sporophores or to the
size and shape of the spores. Drechsler (1919)
was the first to make a detailed study of the
morphology of the actinomycetes that pro-
duce aerial mycelium. Unfortunately, he
limited his study to a small number of cul-
tures; this prevented him from establishing
the existence of many specific types which
could have been recognized on the basis not
only of cultural but also of morphological
properties.
It must be regarded as a considerable step
backward when Lieske (1921) completely
disregarded the work of Krainsky (1914),
Waksman and Curtis (1916), Conn (1917),
and Waksman (1919). He believed that the
classification of actinomycetes was impos-
sible, since the properties observed were
highly variable. His skeptical attitude to-
ward the question of speciation of actino-
mycetes was due largely to his use of complex
media for the growth of these organisms, and
to a lack of sufficient appreciation of the
significance of simple media for their char-
acterization.
8 THE ACTINOMYCETES, Vol. II
Burkholder et al. (1954) were led to con-
clude that the species concepts formulated
by an individual investigator depend a great
deal upon the investigator’s personal expe-
rience, and whether he is a ‘‘splitter’’ or a
‘“Jumper.”’ They suggested further that mi-
crobial species should be characterized by
multiple, readily recognizable, and reason-
ably stable properties; the history of the
cultures and the nature of the medium in
which they are growing are of prime im-
portance.
With the genus Streptomyces gaining con-
siderable economic importance, the creation
of many new species based upon biochemical
properties, notably formation of antibiotics,
resulted in much confusion in the recognition
of some of the species. The use of various
mutagenic agents, such as irradiation, led to
the formation of new forms or strains which
are often markedly different in their nutrient
requirements and biochemical activities from
the mother cultures.
According to this concept, in the classifi-
sation of a group of living organisms, no
single feature can be taken as the predomi-
nant character. Only when this is combined
with a group of other characters is one able
to separate the group into subgroups, no-
tably genera and species. In selecting a char-
acter, no matter what its importance in the
primary subdivision of a group of actino-
mycetes, one may begin with color; or
structure of aerial mycelium; or certain bio-
chemical reactions, which may comprise
proteolytic activities, utilization of carbo-
hydrates, production of antibiotics, or phage
sensitivity. The important thing is to select
a group of properties to characterize each
species, with fewer characters, perhaps only
one, such as antibiotic production, charac-
terizing varieties. One always encounters, of
course, the intermediate forms between the
species. Hach investigator will have to decide
upon the basis of the combination of charac-
ters whether to place an unknown culture
with one species or another. Thus the concept
of species-group or section has come into
being. As a further illustration one may take
S. griseus and S. griseinus, two species be-
longing to the S. griseus group; both are non-
chromogenic; the color of the aerial myce-
hum of both is similar; they are both similar
morphologically; yet they are different from
the standpoint of carbon utilization, phage
sensitivity, and antibiotic production.
Flaig and Kutzner (1954), Kutzner (1956),
Baldacei (1959), and numerous others em-
phasized both physiological and morphologi-
cal criteria. Gause et al. (1957) emphasized
the color of substrate and of aerial mycelium
as well as morphology of sporulating hyphae.
Numerous new species and varieties were
described, although very few prior named
species were discussed or placed into their
system of classification. Many of these spe-
cies and varieties are no doubt synonymous
with previously described forms.
With streptomycetes, the species are
linked together so gradually that it is very
difficult to say where one species ends and
another begins. The creation of ‘‘sections,”’
“oroups,” or ‘‘series’”’ to occupy an inter-
mediate place between genera and species
may help in clarifying relationships, but it
does not do away entirely with the poten-
tial confusion in the creation of new species,
especially when the relation of such species
to those already established is not sufficiently
understood. This confusion has led some in-
vestigators to question ‘‘whether the species
‘
concept is tenable in microbiology, and if it
is not, what we are to substitute for it.” It
has even been suggested that the idea of
static species must be abandoned in favor of
something more elastic.
Even now, after many additional data
have accumulated concerning the morphol-
ogy of the actinomycetes, and after these
organisms have been separated into a num-
ber of genera, there is still no general agree-
ment concerning characterization of species.
SPECIES CONCEPT IN RELATION TO ACTINOMYCETES 9
Krassilnikov (1949) insisted that the shape of
the spore, as seen in the light microscope,
should be recognized as the major criterion
for species differentiation. It is doubtful,
however, whether Krassilnikov’s
“Jongisporus” and “‘globisporus” types, with
their many subtypes, can greatly facilitate
the solution of the problem of species char-
acterization. The cultural properties of these
various
organisms still offer some of the most im-
portant criteria for species differentiation.
There is also now available sufficient addi-
tional information concerning morphology,
such as formation and branching of the
sporophores, formation and nature of spores,
and especially the spore surface as shown by
the electron microscope, to make possible
the use of these criteria not only for supple-
mentary but often for major characteriza-
tion of the species.
Several factors have thus contributed to
the confusion in establishing and recognizing
species of actinomycetes: (a) lack of clearly
defined morphological characters; (b) great
variability of these organisms; (¢) occurrence
of numerous transition types; (d) ease of
formation of mutants; (e) lack of sufficiently
recognizable type species; (f) lack of empha-
sis upon species-groups and upon type cul-
tures; and (g) insufficient recognition of the
formation of well-defined chemical
pounds which could be used as additional
cOom-
criteria for species characterization.
The suggestion that closely related spe-
cies be placed in ‘‘species-groups”’ or ‘‘aggre-
gate-species”’ has recently been gaining con-
siderable attention. Such a unit should be
characterized by various reproducible prop-
erties under standard conditions of culture.
Baldacci et al. (1953, 1956) suggested that
micromorphological criteria, namely, seg-
mentation and branching of vegetative my-
celium, presence or absence of spores, and
arrangement of sporophores, be used for
generic classification. The genus Streptomy-
ces was then divided, on the basis of pigmen-
tation of the vegetative and aerial mycelium,
into a number of ‘series,’ each of which
was further subdivided into species. Gause
et al. (1957) made use of the ‘‘series’’ con-
cept and created a number of groups based
on the pigmentation of the aerial mycelium.
When so many different cultures of ac-
tinomycetes can be isolated easily from natu-
ral substrates, it is but natural that various
intermediate types should be found and that
established species should tend to overlap
one another. If one were to isolate only a
small number of cultures, it would be simple
to recognize a few well defined species. But
when hundreds of similar strains are found
in nature and when many of them show only
minor variations from one another, varia-
tions which are not important enough to
warrant creation of new species but are
nevertheless variations from the established
type, the difficulties mount rapidly (Fig. 2).
When study is based upon a single strain,
a particular species may be described as hav-
ing a yellow or yellowish aerial mycelium.
Another strain may produce, on the same
medium, an aerial mycelium only a shade
different in color from the original type;
this pigment may be designated as sulfur-
yellow, cream-yellow, saffron-yellow, or even
brownish, all other physiological and mor--
phological properties being similar. Would
one be justified in calling such a new strain a
different species? The answer is definitely
“no.” One culture may produce a strong
tyrosinase reaction, and another only a weak
reaction, as indicated by pigmentation with
potato, gelatin, and other protein media.
One would be inclined to accept these as
mere quantitative variations allowable for an
established species. This must be recognized,
since it is well known that had the test been
repeated in another laboratory, where the
medium might be slightly different in com-
position, the method of sterilization of the
medium different, or the age and origin of
the inoculum different,
these variations
LO THE ACTINOMYCETES, Vol. II
Uy
Wey
:
(7
es
te A
\ |
i
Fiaure 2. Schematic representation of tuft and cluster formation by certain Streptomyces species
(Reproduced from: Shinobu, R. Mem. Osaka Univ. Lib. Arts and Ed. B. Nat. Sei. 7, 1958).
might have been sufficient to account for the
minor differences in the color of mycelium
or in the pigmentation of the medium. But
what is one to do when the original culture
is recorded as producing a yellow aerial my-
celium on a given medium, whereas the new
isolate gives a buff or brown mycelium? The
answer would be that if all the other recog-
nizable properties are the same or similar,
this would be nothing more than a variant.
Were one to plate out a single culture and
pick a large number of colonies, similar varia-
tions could no doubt be observed.
Unfortunately, it has frequently been
found much easier to assign undue impor-
tance to these variations and designate a
freshly isolated culture as a new species.
Some justification for this attitude has been
found in the fact that the new culture may
possess an important economic property,
such as the production of a new antibiotic.
It is largely for this reason that within the
last 15 years more ‘‘new”’ species have been
created than in all the previous 75 years
since Ferdinand Cohn first deseribed his
Streptothrix.
Requirements for Adequate Species De-
scriptions
In accordance with the rules of the Inter-
national Code of Nomenclature of Bacteria
and Viruses, certain procedures must be fol-
lowed in describing bacterial species. These
are summarized by Ainsworth and Cowan
(1954) as follows:
The name must be effectively published.
The name must be validated by a concise de-
scription of the diagnostic features of the new
isolate.
The etymology of the name should be ex-
plained.
No Latin diagnosis is required.
When descriptions are reported in a language
unfamiliar to the majority of workers, it is recom-
mended that the authors simultaneously publish
the diagnosis in a more familiar language.
Subcultures of the type strains should be de-
posited at one or more of the national culture
collections.
Unfortunately, these simple rules have
SPECIES CONCEPT IN RELATION TO ACTINOMYCETES 11
not always been adhered to. Numerous
names of actinomycetes are reported in the
literature with no descriptions whatever.
Some of the descriptions have been published
partly in languages not generally accessible,
or in the form of patents, or even as news
announcements in trade or popular journals.
Although every effort has been made in
this treatise to include all species that have
been adequately described, numerous forms
must be listed as “incompletely described”
(Chapter 13). Various names are listed for
which not even an inadequate description is
available, the temptation to name a culture
as a new organism, in order to claim the
discovery, being too great.
Chapter
)
~_
The Genus Actinomyces
The genus Actinomyces comprises anaero-
bie or microaerophilic organisms. They are
mostly pathogenic in nature. The pathogenic
forms are nonacid-fast, nonproteolytic, and
nondiastatic. These have been isolated from
granules in the pus of morbid tissues of a
human and animal disease known as actino-
mycosis. They produce no filterable stages
and show no serological reactions with other
genera.
There are also on record observations con-
cerning the occurrence in various natural
of nonpathogenic, mesophilic,
anaerobic actinomycetes that can with full
justification be included in this genus. Al-
though few of these have been sufficiently
studied, one such species is included. The
saprophytic forms may be proteolytic, ac-
tively fermentative, and may possess marked
substrates
reducing properties.
The natural relationship of this genus to
the other genera of the actinomycetes, based
primarily upon morphological and cytologi-
cal studies, has recently been examined by
Bisset (1959).
Classification of the Genus Actinomyces
I. Pathogenic forms or forms isolated from
pathogenic specimens.
1. Colonies soft, smooth, uniform, not adher-
ent to the medium. No aerial hyphae.
a. Causative agent of certain animal dis-
eases.
lL. Actinomices bovis
b. Isolated from human saliva and carious
teeth.
8. Actinomyces odontolyticus
2. Colonies tougher in texture and warted in
12
appearance, adherent to medium. Aeria
hyphae rare.
a. Hyphae gram-positive and stain faintly
with hemotoxylin. Causative agent of
certain human diseases known as actino-
mycosis.
6. Actinomyces tsraelii
al. Related form.
4. Actinomyces discofoliatus
Hyphae in pus granules stain with basic
stains. Cause of actinomycosis in cats
and dogs.
2. Actinomyces baudetii
Il. Nonpathogenie forms.
1. Occurs in human mouth.
7. Actinomyces naeslundit
a. Related form.
3. Actinomyces cellulitis
2. Occurs in ground waters.
5. Actinomyces hvidhansent
According to Thompson (1950), there are
two distinet species of anaerobic organisms
that should be included in the genus Actino-
myces: A. bovis which is responsible for most
vases of lumpy jaw in cattle, and A. ¢sraeliz
which causes most of the typical infections
in man. This separation of the genus agrees
with the concepts of other investigators.
One strain of A. israeliz was recovered from
a bovine source, and it was suggested that
some bovine infections may be due to A.
tsraeliz. On the other hand, the work of Holm
(1951) and Lentze (1948) indicates that a
small number of human infections may be
due to A. bovis.
Cummins and Harris (1958) fully sup-
ported the conclusions of Erikson (1940)
and Thompson (1950) that bovine and hu-
man strains of Acténomyces are distinct. On
THE GENUS ACTINOMYCES 13
the basis of their chemical data, they sug-
gested that there was very little justification
for placing bovine strains even in the same
genus with the strains of A. zsraeliz. Of the
12 strains received as A. bovis, two were
identical with the human strains, two showed
a cell-wall pattern unlike anything hitherto
recorded, two appeared to be corynebacteria,
and the remaining six formed a homogene-
ous group which seemed to be closely related
to lactobacilli. If cell-wall composition is to
be considered as any guide to the classifica-
tion of these strains, the criteria used for the
identification of A. bovis are insufficient and
many of the investigators who identified the
strain were not properly qualified to do so.
Thompson and Lovestedt (1951) isolated
cultures from the mouths of 24 patients. In
addition to two positive cultures of A.
israeliz, nine of the cultures comprised an
organism which grew under both aerobic and
anaerobic conditions. They considered the
latter to be a saprophyte found in the mouth,
frequently confused with A. The
name A. naeslundii was proposed for these
cultures.
Howell et al. (1959) made a comparison of
200 strains of Actinomyces isolated from the
rsraelit.
oral cavity in the absence of actinomycosis,
and 11 isolated from actinomycotic lesions.
These strains were of two main types, one
corresponding to the organisms described
under the name A. naeslundi7, and the other
essentially identical to those isolated from
lesions, which should be designated as A.
israelit. They recommended that A. naes-
lundit Thompson and Lovestedt be accepted
as the proper name for the rapidly growing
facultative type of Actiénomyces.
One may finally report the results of a
comparative study (Pine et al., 1960) of 11
bovine strains of Actinomyces isolated from
typical cases of lumpy jaw and 15 human
strains which had identified as A.
and A. naeslundi. Of the
strains, one was a typical A. zsraeliz, whereas
been
rsraeliz bovine
the remaining strains formed a homogeneous
group of fast catalase-negative
diphtheroids which invariably failed to form
erowing,
a true mycelium im vitro; they were thus
different from both A.
The last
classical A. bovis. They produced two kinds
israelii and A.
10 strains comprised the
naes-
lundii.
of colonies, depending on the medium: one
smooth colony, identical to that of Coryne-
bacterium acnes, and one rough similar to
that of A. zsraelia but with no mycelium.
They were anaerobes, forming acid from glu-
cose but none from xylose, raffinose, or man-
nitol; nitrates were not reduced and starch
was rapidly hydrolyzed. They were less path-
ogenic for animals than human strains, but
induced lesions in which actinomycotic my-
celial clumps were formed. The A.
strains were also anaerobes; they formed
rsraelar
acid from glucose, usually from xylose and
mannitol, and less often from raffinose; ni-
trates were sometimes reduced to nitrites,
and starch was poorly hydrolyzed if at all.
A. naeslundii strains were facultative anaer-
obes and formed acid from glucose and
raffinose, but none from xylose or mannitol;
nitrates were reduced to nitrites and starch
was poorly hydrolyzed. Micromanipulative
methods for the study of microaerophilic
organisms have been examined by Erikson
(1954); the catalase reaction of A. bovis was
reported by Suter (1956).
According to Emmons,* there is little
value in presenting as valid all the following
species until they have been studied carefully
in pure culture. He suggested to accept only
A. bovis, A. israelii, A.
*
baudetii, and A.
naeslundi. He went so far as to suggest that
the staining reactions of A.
hardly sufficient for its differentiation.
baudetiz are
Descriptions of Species of Actinomyces
1. Actinomyces bovis Harz (Harz, C. O.
In Bollinger, O. Centr. med. Wiss. 15: 485,
* Personal communication.
14 THE ACTINOMYCETES, Vol. II
Figure 3. A. bovis, branching mycelium; cul-
tured from human tonsils, X 1000 (Reproduced
from: Emmons, C. W. Puerto Rico J. Public
Health Trop. Med. 11: 720, 1936).
1877; Jahr. Miinch. Thierarzeneisch 5: 125,
LSA):
Actinomyces bovis was the first authentic
actinomycete described as a causative agent
of disease; it is natural, therefore, that it
should have a number of synonyms. These
are given here, without any guarantee that
the list is complete.
Synonyms: Discomyces bovis Rivolta,
1878; Bacterium actinocladothrix Afanasiev,
1888; Nocardia actinomyces de Toni and Tre-
visan, 1889; Actinomyces hominis Bostroem,
1890; Streptothrix actinomyces Rossi-Doria,
1891; Cladothrix bovis Macé, 1891; Oospora
bovis Sauvageau and Radais, 1892; Actino-
myces albidoflavus Rossi-Doria, 1891; Actino-
myces sulphureus Gasperim, 1894; Nocardia
bovis R. Blanchard, 1895; Streptothrix israeli
Ixruse, 1896; Cladothrix actinomyces Macé,
1897; Streptothrix actinomycotica Foulerton,
1899; Discomyces bovis R. Blanchard, 1900;
Streptothrix spitzi Ligniéres, 1903; Sphaeroti-
lus bovis Engler, 1907; Cohnistreptothrix
israeli Pinoy, 1911; Actinomyces israeli Vuil-
lemin, 1931. See also Baldacei (1937).
Morphology: Grows in the form of sulfur-
colored granules in the pus of cases of ac-
tinomycosis. The radiating hyphae are cov-
ered with extraneous material deposited by
the host to form clubs. Organism is gram-
positive, nonmotile, nonacid-fast. Colonies
are dull white in color, only slightly ad-
herent to the medium. No aerial hyphae.
Mycelium undergoes fragmentation very
rapidly into V- and Y-forms. Extensive
branching is rare. Hyphae less than 1 » in
diameter (Fig. 3).
Semisolid media: Growth excellent, espe-
cially with paraffin seal. No soluble pigment
produced.
Gelatin: Growth secant, flaky. No lique-
faction.
Liquid media: Occasional turbidity with a
light, floceulent growth.
Egg or serum media: No proteolytic ac-
tion.
Milk: Turns acid; no coagulation and no
peptonization. Sometimes there is no growth.
Sugar utilization: Acid from glucose, su-
crose, and maltose; no acid from salicin or
mannitol.
Temperature: Optimum 37°C. Does not
grow at 22°C. Killed at 60°C.
Oxygen requirement: Anaerobic to micro-
aerophilic. Grows readily in an atmosphere
of CO,.. Bovine strains are more oxygen-
tolerant on egg or serum media than strains
of human origin.
Viability: Pure cultures do not live more
than 10 to 14 days. On Dorset’s egg medium,
they may survive in an ice chest for 3 to 4
weeks.
Habitat: Originally found in lumpy jaw of
THE GENUS ACTINOMYCES 15
cattle. Usually found in and about mouths of
animals.
Remarks: King and Meyer (1957) re-
cently suggested that in order to implement
proper identification of A. bovis, certain se-
lected differential criteria, such as catalase
test, litmus milk reactions, and the utiliza-
tion of xylose, salicin, and raffinose, can be
used. Slack and Moore (1960) suggested the
use of fluorescent antibody formation for the
further identification of this organism.
2. Actinomyces baudeti Brion, 1942 (Brion,
G. de. Rev. de Méd. Vétér. 91: 157, 1942;
Brion, G. de, Goret, and Joubert. Proc. VI
Congr. Intern. Patol. Comp., Madrid 1: 48,
1952).
Morphology: Granules from histological
preparations show tangled, radiating hy-
phae; ends of hyphae rounded and ovoid,
forming a crown. Hyphae take basic stains.
Mycelium composed of slender hyphae, 0.2
to 0.4 pw. Nonseptate. Ends swollen and
rounded. Copious branching. In artificial
media hyphae are frequently short, rarely
exceeding 20 uw in length.
Agar colonies: Dull, whitish granules ad-
hering slightly to the medium.
Liquid media: A sediment of white gran-
ules is produced.
Gelatin: No liquefaction.
Blood serum: In 4 to 5 days, surface cov-
ered with white granules which are the size
of a pin head.
Serum media: No proteolytic action.
Brain extract: Growth favored in some
media.
Indol: Production slight.
Sugar utilization: Acid from glucose, su-
crose, and starch.
Oxygen demand: Anaerobic to microaero-
philic.
Optimum temperature: 37°C.
Pathogenicity: Pathogenic when inocu-
lated into dogs, rabbits, and guinea pigs
(forms subcutaneous abscesses).
Source: Isolated from various types of le-
sions in cats and dogs.
3. Actinomyces cellulitis (Linhard, 1949)
nov. comb. (Linhard, J. Ann. inst. Pasteur
76: 478, 1949).
Synonym: Actinobacteriwm cellulitis Lin-
hard.
Morphology: Polymorphic rods, showing
primary, secondary, and sometimes tertiary
branching. Length 5 to 7 uw, diameter 0.6 wu.
Nonmotile. Gram-positive.
Agar media: Colonies lenticular. No gas.
Glucose broth No. turbidity.
Abundant growth, settling to bottom.
Gelatin: No liquefaction.
Milk: Unchanged.
Serum: Serophilic, but can be adapted to
serum-free media.
Nitrate reduction: Positive.
Oxygen demand: Anaerobic and micro-
cultures:
aerophilic. Colonies produced at 4 to 5-mm
depth in agar media.
Reduction: Does not reduce neutral red or
safranin.
Carbon utilization: Positive utilization of
glucose, fructose, maltose, galactose, and
sucrose. Produces volatile acids (propionic
and formic). Production of gas may suggest
either a contaminated culture or the absence
of an Actinomyces.
Pathogenicity: Nonpathogenic.
Habitat: Oral cavity of man.
4. Actinomyces discofoliatus — (Griiter,
1932) Negroni (Negroni, P. Mycopathol. 1:
81-87, 1938-1939).
Morphology: Deep colonies in semisolid
glucose agar are whitish, lens-shaped, crossed
or forming dihedral angles; margins of colo-
nies regular; consistency of colonies slimy.
Bacteria-like entities measuring 3 to 4 u to
10 to 15 w by O.8 yw, occurring as isolated
elements or V- or Y-shaped elements. Com-
pact colonies in hanging-drop cultures. The
filaments have a tendency to dichotomous
16 THE ACTINOMYCETES, Vol. II
FicurReE 4. A.
Institute of Pathology).
branching, with prevailing development of
one branch.
Glucose agar: Discoid, moist, and brilliant
colomes; shghtly elevated in the central part
with nearly regular margins.
Gelatin: No liquefaction.
Glucose broth: Slimy sediment and some-
times a shght turbidity. The medium be-
comes clear at the end of 8 to 10 days.
Carbon sources: Acid but no gas from glu-
cose, maltose, fructose, lactose, sucrose, and
inulin; very little or no acid from mannitol.
Starch: Not attacked.
Sucrose: Not inverted.
Nitrate reduction: Negative.
H.S: Formed.
Indol: Shght quantity produced.
Fats: Shghtly attacked.
Olive oil: Not attacked.
Optimum temperature: 37°C.
Oxygen demand: Facultatively anaerobic.
Remarks: Vitality weak. Deep cultures in
semisolid media die if held for longer than 8
le dey wal
to 10 days at 37°C, or for longer than 30
minutes at 60°C. Exposure for longer than
israelii, grown anaerobically in veal infusion agar, X 975 (Courtesy of Armed Forces
a few minutes in dilute mineral acids kills
the organism. The organism can be kept
alive for 2 to 3 months if cultures are kept
in an ice chest, in a dried state, or under
vacuum.
Habitat: Lachrymal concretions and hu-
man actinomycotic lesions.
5. Actinomyces hvidhanseni (Hvid-Han-
sen, 1951) nov. comb. (Hvid-Hansen, N.
Acta Pathol. Microbiol. Scand. 29: 335-338,
1951).
Synonym: Actinomyces israeli Hyid-Han-
sen.
Morphology: Gram-positive, nonacid-fast,
nonmotile. Polymorphic, bent, and often
branched rods. Obligately anaerobic.
Meat liver agar: Colonies circular or ir-
regular, often in the form of bodies bounded
by four concave surfaces meeting in four
acute vertices, of highly varying size and of
a pale pink color. Surface colonies circular,
convex, grayish-white or white; transparent
S-colonies of a butyrous, viscous, but not
mucous consistency.
Meat liver broth: Diffuse growth at first,
THE GENUS ACTINOMYCES 17
followed rapidly by a fairly voluminous pale
pink, homogenous precipitate.
Thioglycollate medium: Growth either
diffuse, netlike, or dispersed and granular.
Gelatin: Liquefied.
Milk: Coagulated in 24 to 48 hours and
peptonized in 3 weeks.
Blood: All strains hemolyze human blood
on solid media but do not form a soluble
hemolysin.
Sugar utilization: Galactose, fructose, and
glycerol vigorous; inulin, maltose, mannitol,
starch, duleitol, and
somewhat less readily; xylose and arabinose
not at all.
Reduction: Some strains form a little hy-
drogen sulfide. Sulfites and sulfates are not
reduced. Nitrates reduced to nitrites and in
saccharose, lactose
some cases to ammonia. Safranin, phenosaf-
ranin, or neutral red reduced. Most
strains decolorize methylene blue in 4 to 24
not
hours; some do not.
Temperature: Optimum 37°C. Heating to
50-60°C for 15 minutes injurious.
Remarks: All strains catalase-positive. All
produce ethyl alcohol, aldehyde, acetone,
ammonia. A faint indol reaction is found in
alkaline distillate. The presence of volatile
acids, tartaric acid, and lactic acid has been
demonstrated, but not succinic acid. Pro-
plonic acid and formic acid in ratios of from
3 to 1 up to 20 to | for the six strains ex-
amined.
Habitat: Ground water.
Remarks: Kalakoutski (1960) found an-
aerobic actinomycetes in natural waters and
in the air of apartments occupied by man,
but not in the soil.
6. Actinomyces israelii (Kruse) Lachner-
Sandoval, 1898 (Wolff, M. and Israel, J.
Arch. pathol. Anat. 126: 11, 1891).
Synonyms: Streptothrix
1896;
Actinomyces bovis Wright,
Prumpt, 1906;
israeli Sampietro, 1908;
Kruse,
Gedoelst, 1902;
1905; Discomy-
rsraeli
Discomyces israelr
ces bovis Actinobacterium
Cohnistreptothria
israeli Pinoy, 1913; Nocardia israeli Castel-
lani and Chalmers, 1913; Brevistreptothrix
israeli Ligniéres, 1924; Proactinomyces israeli
Jensen, 1931; Corynebacterium israeli Lentze,
1938; Actinomyces israeli var. indo-sinensis
Reynes, 1947.
Morphology: Large, club-shaped forms
are seen in morbid tissues. Substrate myce-
lium consists of rapidly septating and spor-
ulating hyphae. The branches may extend
into the medium in long filaments or may
exhibit fragmentation and characteristic an-
gular branching. Hyphae occasionally sep-
tate, but no definite spores are formed.
Colonies exhibit a considerable degree ot
polymorphism, but no stable variants have
been established. Colonies are tougher in
texture than those of A. bov7s. Old colonies
warted in appearance (lig. 4).
Gelatin: Growth scant, flaky. No liquefae-
tion.
Liquid media: Growth in form of white
compact colonies or granular sediment. Me-
dium shows no turbidity, usually remaining
clear. No gas and no odor.
Pigments: No soluble or insoluble pig-
ments.
Ege media: No proteolytic action.
Milk: but
not clot. No peptonization. Frequently no
Becomes acid, usually does
growth.
Starch: Shght hydrolysis.
Oxygen requirement: Anaerobic.
Nitrate reduction: Generally negative.
Sugar utilization: Greater ability to utilize
sugars than A. bovis. Acid but no gas from
glucose, galactose, lactose, fructose, maltose,
raffinose, sucrose; no acid from inulin.
Hemolysis: Slight to marked.
Serological reactions: Lack of serological
affinity with A. bovis.
Temperature: Optimum 37°C. Destroyed
at 55-60°C in 30 minutes.
Habitat: Dental caries, tonsils, and natu-
ral cavities of man and animals. Chief etio-
logical agent of human actinomycosis, de-
Re) THE ACTINOMYCETES,
J
oe’
Figure 5. A. israelii (Reproduced from: Pré-
Vota Au RR tas Intern. Congr. Microbiol., Symp.
Actinomycetales, Rome, 1953, p. 45).
scribed first by Wolff and Israel (1891) and
later by Wright (1905).
temarks: Vitality weak. Cultures no
longer viable after 8 to 10 days. Erikson and
Porteous (1953) succeeded in obtaining good
growth by continued subculture in a medium
containing 99 parts of 1 per cent casein hy-
drolyzate and 1 part of heart broth and 0.5
per cent glucose. Antigenic structure of or-
ganism has been recently studied by Itwa-
pinski (1960).
According to Grootten (1934)
ism is highly polymorphic. Rods varying in
length are formed in young culture. They are
straight or slightly curved, with round
oval extremities. Occasionally, long or even
filamentous forms are found. Some of the
filaments end in small spherical or pear-
shaped swellings. It does not form spores.
t does not grow in the upper
5- to 10-mm zone; below that zone, it forms
a layer of 2 to 4 mm with numerous small
in the deeper layers, the colonies
attain diameters of 2 to 3
, the organ-
In agar tubes, 1
colonies;
are fewer,
No
Liquid media remain clear.
nonproteolytic; milk is not coagulated. Blood
but may
mm. gas and no odor are produced.
The organism is
Vol. II
is rapidly hemolyzed. It does not grow on
potato plugs, except poorly when glycerin-
ated. It slowly attacks glucose, lactose, mal-
tose, sucrose, and mannitol. It does not grow
in glucose-gelatin medium. Animal infection
is obtained by introducing a culture into the
peritoneum of rabbits.
Negroni (1954) deseribed <A.
further detail.
israelit in
Deep colonies in semisolid
glucose agar are globous, 1 to 2 mm in
diameter, whitish, opaque, and with an ir-
regular surface. Colonies are of a
consistency and cannot be homogeneously
suspended in water. On glucose or glycerol
agar slants, the colonies are elevated, mam-
cheesy
milated, and whitish, with moist and_ bril-
liant surface and irregular margins. Sub-
merged mycelium is well developed. The
colonies have a cheesy consistency and can
sasily be removed from the medium with a
platinum loop (Fig. 5).
According to Erikson and Porteous (1955),
the conversion of a “rough” typical strain of
A. israelii to a ‘“‘smooth” soft form more
tolerant of oxygen is a result of the physical
trapping within the mycelium of a few alien
facultative anaerobes, usually staphylococci.
Actinomyces naeslundit Thompson and
Lovestedt, 1951 (Thompson, L. and Love-
stedt, S. A. Proc. Staff Meet. Mayo Clinic
26: 169, 1951).
Morphology: Organism forms small, whit-
ish, firm colonies. Mycelial branching, but no
segmentation. Not acid-fast.
Artificial media: Good growth.
Hormone agar: Rough and smooth colo-
nies, | to 2 mm in diameter, after 4 days.
Surface of colonies varies from smooth to
nodular to wrinkled. Consistency varies from
butyraceous to tough and adherent. Colo-
nies are opaque, with color varying from
white to cream.
Glucose brain broth: Growth rapid and
abundant. Acid produced.
Gelatin: Growth slow. No liquefaction.
Starch: Not hydrolyzed.
THE GENUS ACTINOMYCES 19
Milk: Growth scant or absent.
Aerobiosis: Grows both under aerobic and
anaerobic conditions, somewhat better aero-
bically.
Temperature: Optimum at 37°C;
growth at 32°C.
Pathogenicity: Nonpathogenic.
Habitat: Human mouth. Considered to be
a saprophyte found in the mouth and fre-
quently confused with A. zsraeliz.
some
8. Actinomyces odontolyticus Batty, 1958
(Batty, I. J. Pathol. Bacteriol. 75: 455-459,
1958).
Morphology: At first, the organism ap-
pears in the form of short rods subdivided
by one or two transverse septa. Later, these
rods gradually elongate until a septate sub-
mycelium is produced. At the end of these
filaments, globular ‘“‘initial cells’? are pro-
duced which germinate to produce a non-
septate secondary submycelium, which soon
commences to break up. Finally, in a week
to 10 days, small spores commence to form
singly upon short side branches. The size of
the mature spores varies greatly in different
strains.
Appearance of colonies: Colonies are usu-
ally few in number with an initial appearance
similar to those of a-hemolytic streptococci
of comparable age. Later, they develop a
dark red hemin-like pigment,
tinguishable. At and after this stage the
colonies are exceedingly difficult to subcul-
ture. After prolonged artificial culture the
organisms can be subcultured at any stage.
Attempts to isolate the organism upon horse
serum agar or nutrient agar are usually un-
successful, but after several subcultures a
sasily dis-
profuse growth of small convex nonpig-
mented colonies is obtained on both these
media. All strains grow equally well under
aerobic and conditions at all
stages in their life cycle; in agar stab cultures
a filiform growth is obtained throughout the
line of inoculum. Growth in peptone broth is
sparse, but in this medium enriched with
anaerobic
odontolyticus, various stages of
Ficure 6. A.
culture development (Reproduced from: Batty, I.
J. Pathol. Bacteriol. 75: 455-459, 1958).
yeast extract, a characteristic glutinous ropy
sediment is produced which disperses to
give an even turbidity (Fig. 6).
No strain produces catalase, oxidase, in-
dole, hydrogen sulfide or acetylmethylear-
binol; all are methyl red-negative and all fail
to ferment fructose, trehalose,
starch, inulin, dextrin,
rhamnose, glycerol, dulcitol, and salicin. A
maltose,
glycogen, xylose,
few strains ferment sucrose, galactose, arabi-
nose, or mannitol, with the production of
acid but no gas. About half the strains pro-
duce ammonia from peptone, acidify and
coagulate litmus milk, and are tolerant of a
concentration of 1 in 4000 potassium tellu-
rite. Some strains hydrolyze urea. None
liquefy gelatin, Loeffler’s medium, or coagu-
nitrate to
lated egg medium. All reduce
nitrite within 18 hours.
20 THE ACTINOMYCETHES, Vol. II
Habitat: Human saliva in deep dental
caries.
Remarks: This organism
bovis in its life cyele.
resembles A.
Incompletely Described Forms of Ac-
tinomyces
In addition to the above well described
and readily recognizable forms belonging to
the genus Actinomyces, numerous other an-
aerobic organisms have been listed in the
literature. Some of these organisms are no
doubt strains of the well described forms and
their names would be in synonymy. Others
may represent distinct species.
It is of particular interest to cite, in this
connection, the ideas of Prévot (1957), who
considered Actinomyces bovis as an aerobic
organism and, therefore, the genus Actino-
myces as an aerobic group. He suggested
that Actinomyces israelii represent the an-
aerobic group, and the generic name Actino-
bacterium Haas, 1906 (Syn. Cohnistrepto-
thrix Pinoy, 1913) be given priority for desig-
nating the anaerobic forms. Prévot (1957)
divided the genus Actinobactertum into six
species: (1) A. zsraeliz, (2) A. meyers, (3) A.
abscessus, (4) A. liquefaciens, (5) A. cellu-
litis, and (6) A. propionict.
In addition to the above, numerous other
anaerobic forms have been described, such
as A. canis Levy, 1899; A. interproximalis
Fennel, 1918; and others. Some additional
names will be found in Chapter 13.
=)
Chapter 3
The Genus Nocardia
Characterization of Genus
The genus Nocardia represents a
of aerobic actinomycetes which includes both
pathogens and saprophytes. The relation-
ship of this genus to, and possible overlap-
ping with, the genus Mycobacterium, on the
one hand, and the genus Streptomyces, on the
other, have already been discussed (Volume
I). Numerous cultures of nocardiae have
been isolated from human and animal infec-
tions, and claimed to be the causative agents
of the particular disease. The fact, however,
that a culture of an organism has been iso-
lated from a lesion of a man or an animal is
no proof that it is primarily responsible for
the particular disease; it may actually be a
secondary invader or a member of a mixed
infection. Some species of Nocardia are def-
group
initely associated, however, with certain
diseases, or have at least been isolated from
infected tissues. This gave origin to the term
“nocardiosis,’ descriptive of these disease
conditions.
The colonies produced by nocardiae are
either smooth, or rough and much folded;
they are either of a soft or dough-like con-
sistency, or compact and leathery, especially
in early stages of growth. Many species of
Nocardia do not form any aerial mycelium;
some give rise to a limited aerial mycelium
which may structurally be similar to that of
the substrate mycelium; still others may
produce aerial hyphae and spores which may
be indistinguishable from those of Strepto-
myces and are thus responsible for various
cases of overlapping between these two
genera.
Nocardias multiply by concentration
and segmentation of the protoplasm within
a filamentous cell, followed by dissolution of
the cell membrane. The fragmented portions
of the mycelium usually develop into fresh
mycelium under favorable conditions, either
by germ tubes or by lateral budding. Strep-
tomycetes produce true spores or conidia, the
vegetative mycelium not segmenting spon-
taneously into bacillary or coecoid forms, but
remaining nonseptate and coherent even in
old cultures, thus producing the characteris-
tic tough textured, leathery growth. In no-
cardiae, the aerial mycelium represents an
extension upward of the vegetative myce-
lium; it does not exhibit any differentiated
protoplasm and is sterile and abortive. When
a streptomycete has lost the capacity of
producing aerial mycelium, a form analogous
to that of a nocardia may result, except for
the structure the mycelium and the
capacity of the degenerated streptomycete
to regain the lost capacity. It is occasionally,
therefore, a matter of personal preference
whether to place a freshly isolated culture in
of
one genus or another. Some nocardiae are
acid-fast or partially acid-fast, and others
are not.
The mode of branching of the substrate
mycelium (see Volume I, Chapter 5), the
biochemical properties (proteolytic and sero-
logical activities), and chemical nature of the
cell walls of nocardiae appear to distinguish
them from the streptomycetes. Hoare and
22 THE ACTINOMYCETES, Vol. II
Work (1957) have shown that these genera
‘an be differentiated by the configuration of
the diaminopimelic acid present in whole cell
hydrolysates; streptomycete cell walls con-
tain the L-isomer, whereas nocardiae cell
walls contain the pL-isomer. Cummins and
Harris (1958) reported that the presence or
absence of arabinose in the hydrolysates of
the intact organisms can also be used to
identify them; nocardiae cell walls contain
arabinose, whereas streptomycete cell walls
do not. The sensitivity of most Streptomyces
species, but not of nocardiae, to the action of
lysozyme on their cell wall preparations,
studied by Sohler, Romano, and Nickerson
(Volume I, p. 159), provides further criteria
for distinguishing between members of these
two genera. Studies of infrared absorption as
a taxonomic criterion (Riddle et al., 1956)
has also been suggested.*
In view of the overlapping between certain
forms placed for convenience in either one
genus or the other, the separation of atypical
strains of Nocardia or Streptomyces by mor-
phology or fermentation tests alone may be
difficult, as pointed out by Gordon and
Mihm (1957).
The genus Nocardia has been described in
the last edition of Bergey’s Manual as
follows:
“Slender filaments or rods, frequently
swollen and occasionally branched, forming
a mycelium which, after reaching a certain
size, assumes the appearance of bacterium-
like growths. Shorter rods and coccoid forms
are found in older cultures. Conidia not
formed. Stain readily, occasionally showing ¢
slight degree of acid-fastness. Nonmotile.t
* Personal communication from Dr. N. M.
McClung.
7+ The existence of motility among the nocardias
was considered by Jensen (1953) as indisputable,
and this really is not surprising in view of the
numerous observations on motility in the closely
related coryneform bacteria. The species in the
order Actinomycetales cannot any longer be re-
garded as constantly nonmotile (Fig. 14).
No endospores. Aerobic. Gram-positive. The
colonies are similar in gross appearance to
those of the genus J/ycobacterium. Paraffin,
phenol and m-cresol are frequently utilized
as a source of energy.
“In their early stages of growth on culture
media (liquid or solid), the structure of
nocardias is similar to that of actinomycetes
in that they form a typical mycelium; hy-
phae branch abundantly, the branching
being true. The diameters of the hyphae vary
between 0.5 and 1 yu, usually 0.7 to 0.8 pn,
according to the species. The mycelium is
not septate. However, the further develop-
ment of nocardias differs sharply from that
of actinomycetes; the filaments soon form
transverse walls and the whole mycelium
breaks up into regularly cylindrical short
cells, then into coecoid cells. On fresh culture
media, the coccoid cells germinate into my-
celia. The whole cycle in the development of
noecardias continues for 2 to 7 days. Most
frequently the coccoid cells are formed on
the third to fifth day, but in certain species
they can be found on the second day.
“The multiplication of nocardias proceeds
by fission and budding; occasionally they
form special spores. Budding occurs often.
The buds are formed on the lateral surface
of the cells; when they have reached a cer-
tain size, they fall off and develop into rod-
shaped cells or filaments. The spores are
formed by the breaking up of the cell plasm
into separate portions usually forming 3 to
5 spores; every portion becomes rounded,
covered with a membrane and is transformed
into a spore; the membrane of the mother
cell dissolves and disappears. The spores
germinate in the same way as those of ac-
tinomyces. They form germ tubes which
develop into a mycelium (Fig. 7).
“The colonies of nocardias often have a
paste-like or mealy consistency and can
sasily be taken up with a platinum loop; they
spread on glass and occasionally render the
broth turbid. The surface colonies are
THE GENUS
. Fie
|
»
p :
F as = 4 ‘
‘ 4
ad, ’
4 te. ~~ Lf ’
, 7 ‘ Ps « 4
x \ ‘\ at ea A SG eS
> / 7 Reena
ay ‘ SF 2 a7
{ ~
N ee Ne
aN : oe
ged — ey
“NN
te
Pad
¥ é
*
*
i 4 g.
be + %
4 j
? én
ee e *
— .
* ia & Cc
*
4 : ?
#
Ficure 7. N.
NOCARDIA 23
opaca: (a) grown for 4 days on n-dodecane and mineral salts; gram stain, X 960; (b)
same grown 3 days; X 3700; (c) same as (a) but using fat stain, X 1920; (d) two-day growth, X 12,500
(Reproduced from: Webley, D. M. J. Gen. Microbiol. 11: 425, 1954).
smooth, folded or wrinkled. Typical nocar-
dias never form an aerial mycelium, but
there are cultures whose colonies are covered
with a thin coating of short aerial hyphae
which break up into cylindrical oidiospores. *
Many species of nocardias form pigments;
their colonies are of a blue, violet, red,
yellow or green color; more often the cultures
are colorless. The color of the culture serves
as a stable character. The type species is
Nocardia farcinica Trevisan.”
Classification of Nocardia Species
De Toni and Trevisan (1889) described
five species of Nocardia: N. farcinica, N.
actinomyces, N. foerstert, N. arborescens, and
N. ferruginea.
* See the work of Gordon and Mihm (1958).
Jensen (1932a) found that a number of
organisms previously described as species of
Mycobacterium actually belong, on account
of definite mycelial growth in the initial
stages of their life cycles, to the genus Nocar-
dia.* Mycobacterium agreste Gray and Thorn-
ton and B. mycoides corallinus Hefferan were
found to be similar to one another and were
The
same was true of JJ. salmonicolor den Dooren
regarded as one species, N. corallina.
de Jong, which was designated as N.. sal-
monicolor. Mycobacterium opacum den
Dooren de Jong and JM. crystallophagum
Gray and Thornton proved to be identical
and were named N. opaca. Mycobacterium
erythropolis, a closely related form, was des-
* Generic name Proactinomyces used.
Ficure 8. N. paraffinae, showing a section of a
colony covered with mature aerial mycelium (Re-
produced from: Hirsch, P. and Engel, H. Ber.
Deut. Botan. Ges. 69: 454, 1956).
Fiacure 9. Nocardia, strain 70, showing the de-
velopment of the aerial mycelium on mineral agar
(Reproduced from: Hirsch, P. and Engel, H. Ber.
Deut. Botan. Ges. 69: 454, 1956).
LI;
THE ACTINOMYCETES, Vol. II
A. Unstable mycelium (a-form), with
short mycelium (if formed at all),
bacterial (diffuse) growth in liquid
media, bacteria-like colony.
Stable mycelium (8-form), producing
long hyphae, colony growth in liquid
media, and Streptomyces-like type of
colony.
Umbreit (1939) modified the system of
B.
Jensen as follows:
I. Partially acid-fast, nonproteolytic, non-
diastatic; constantly utilize paraffin.
1. a-Mycelium type: N. opaca,* N. eryth-
ropolts.
. B-Mycelium type:
a. Red-colored: N. polychromogena
N. asteroides.
b. Yellow-colored: NV. paraffinae.
Nonacid-fast forms, constantly diastatic
1. a-Mycelium type:
a. Nonproteolytic: N. mesenterica.
b. Proteolytic: N. actinomorpha.
2. B-Mycelium type:
a. Yellow-colored: N. flavescens.
b. Red to orange: N. maculata.
Krassiinikov (1938) divided the genus
Nocardia* into two groups:
re)
I. Well developed aerial mycelium, with
ignated as N. erythropolis. Microbacterium
mesentericum Orla-Jensen was renamed N.
mesentericus.
Jensen divided the genus Nocardia into
two distinct groups:
I. Nonproteolytic organisms with strongly
refractive cells showing a partial acid-
fastness in milk and sometimes in other
media; capable of decomposing paraffin.
Some species of this group form a transi-
tion to the genus A/ycobacterium.
II. Mostly proteolytic forms with weakly
refractive, nonacid-fast cells. This group
forms a close transition to the forms now
included in the genus Streptomyces.
A further separation of the genus was
based upon the structure of the aerial myce-
lium.
Ale
substrate mycelium seldom producing
cross walls. The hyphae break up into
long, thread-like rods. Branches of the
aerial mycelium produce segmentation
spores and oidiospores; the latter are eyl-
indrical with sharp ends. No spirals or
fruiting branches. This is the same as
group 8 of Jensen.
Typical nocardial forms. Mycelium de-
velops only at early stages of growth,
then breaks up into rod-shaped and
coccoid bodies. Smooth and rough col-
onies, dough-like consistency, similar to
bacterial colonies. Aerial mycelium not
formed or only around colonies.
* Generic name Proactinomyces used.
THE GENUS NOCARDIA 25
TABLE |
Summary of growth characteristics of 18 strains of Nocardia (McClung, 1949)
Fragmentation
Organism Scion |branchiae | branching
Age Type 1 Type 2 Type 3
hr hy hr hr q % %
Group I
WieBie ae SRL We le Sach toh 2 8 16 0 13 70 30 0
INESOORLOSUUS Hera ee Stef 6 12 0 12 60 30 10
SU bo ial ene ea 9 15 0 13 50 50 0
BB eeeaer ete tees er 10 36 0) 14 50 50 0
N. erythropolis...... ; 7 1] 14 0 14 69 40) 0
N. polychromogenes... ‘rE 1] 13 0 14 40 30 30
Group II
NSO Ree eae ec tes Sate 1] 12 5 19 6 28 66
‘oleae SA ae meee 10 12 7 1S 5 40 50
INGETALD Chetan esc 10 14 1 20 5) 30 65
N. polychromogenes............ 14 30 20 120 Unknown
IN@rastenovdes:s.<..)... 10 28 96 96 Unknown
Group III
ees Met ewer Paes ee 529s ah Sache ee oe: 10 16 4 0 0 0
Pedal errant erty t ora cat's loc). -yerantontrs 9 1] 6 0 0 0
tse O08 Sie) fue ae eae a 10 12 S 0) 0 0
NOE Zeta eaten! Staves ne Seetehehs aust 11 15 6 0 0 0
WINS) Shh alk o Aleantsi eo eeee ecere ee 10 15 5) 0 0 0
Hato goa ra Riek Oke i eres ee pate DR a eee 10 13 9 8) 0) 0
2) = OST O IE T TN Eco cous ys chen epes sis anes Bes Ss 13 7 0 0) 0
MeClung (1949) divided the genus Nocar-
dia into three groups:
I. Scant mycelial development, sparse
branching. Colonial texture soft, pasty,
and sometimes mucoid; pigment intra-
cellular and insoluble.
Extensive mycelial development,
straight branches which do not over-
lap. Colonial texture soft and pasty;
pigment intracellular and insoluble.
oxtensive mycelial development,
fragmentation of hyphae, contorted and
profusely produced branches which
overlap. Colonial texture
‘artilaginous; generally both intracel-
lular and soluble pigments are pro-
duced (Figs. 8, 9).
The pattern in Nocardia fragmentation
can be separated into three types. In Type
1, an acute angle is formed in a hypha pre-
ceding division, which occurs at the apex of
II.
III.
no
Waxy or
the bend; following division the new hyphal
tips grow out parallel to each other. In Type
2, division occurs in a straight or slightly
curved portion of a hypha; following di-
vision, the newly formed ends bend slightly
and grow past each other. In Type 3, di-
vision occurs in the parent hypha close to or
at the juncture of a branch; a new hypha
may grow from the place of division at
the base of the branch; the newly formed
hyphal tip bends and continues to grow.
Type 1 fragmentation is characteristic of
Group I, and Type 3 of Group II. Type 2
occurs in both groups, and Group III lacks
fragmentation (Fig. 10).
A summary of the growth characteristics
of various strains of Nocardia belonging to
these three groups is presented in Table 1.
In an attempt to find a group of depend-
able properties for the separation of the
genera Nocardia, Streptomyces, and Mvyco-
26 THE ACTINOMYCETES, Vol. II
Fieure 10. N. rubra: (above) 24 hr, glycerol
nutrient agar, methylene blue; (below) same,
stained with Sudan black B, X 1600 (Reproduced
from: McClung, N. M. Lloydia 12: 165, 1949).
bacterium, Gordon and Smith (1955) exam-
ined 152 cultures labelled Streptomyces and
99 cultures labelled Nocardia; those strains
of the latter that formed soft, fragmenting,
vegetative mycelium were excluded. Of the
cultures designated as Streptomyces, 83 per
cent produced an aerial mycelium typical of
this genus; 13 per cent failed to produce
aerial mycelium, although a few formed rudi-
mentary aerial hyphae; inability to form
spores was considered as a lost property,
the physiological reactions of the
strains were the same as in the previous
since
group of sporulating cultures; only five
strains, or + per cent of the cultures possessed
nocardial properties. Of the cultures desig-
nated as Nocardia, 68 produced aerial hy-
phae, varying from rudimentary to luxuri-
ant, some even forming chains of spores.
According to their physiological properties,
24 of these cultures should have been desig-
nated as Streptomyces. A few of the strains
could be considered as intermediate between
the two genera.
Gordon and Mihm (1957) further reported
the results of an examination of 219 cultures
labelled Streptomyces, 214 Nocardia, and 243
TABLE 2
Certain physiological and biochemical characteris-
tics of various strains of two species of Nocardia
(Gordon and Mihm, 1959)
N. asteroides N. brasiliensis
(98 strains), (50 strains),
Property positive positive
strains strains
Decomposition of :
Casein 0 98
Gelatin 36 100
Tyrosine ) 100
Xanthine 0 0)
Hydrolysis of starch 58 56
Acid from:
Adonitol 0) 0
Arabinose 0 0
Erythritol 6 0
Galactose 24 92
Glucose 100 96
Glycerol 9S 98
Inositol 2 100
Lactose 0 0)
Maltose 6 )
Mannitol ) 94
Mannose 19 68
a-Methyl-p-glucoside ) 0
Raffinose 0) 0
Rhamnose 33 0
Sorbitol 0 0
Xylose ) 0)
Nitrite from nitrate Sd 92
Growth at:
50°C 24 8)
40°C 90, 56
35°C 100 100
28°C 100 100
10°C 12 30
Utilization of:
Acetate 100 100
Citrate 33° 98
Malate 100 100
Propionate 100 100
Pyruvate 100 100
Succinate 100 98
Benzoate 0) 0)
THE GENUS NOCARDIA
TABLE 3
Comparative properties of certain acid-fast nocardias (Suter, 1951)
Aerobic
. fastidiosa
. leishmanii
. caprae
4
. pretoriana
. pulmonalis
. paraffinae
. transvalensis
4
4
baths TE tis
. polychromogenes
4
. minima
4
. coeliaca
. rubropertincta
. asteroides
‘
. salmonicolor
4
rubra
. farcinica
b+++1+++++4+4+4+44 1
4
aie 98> |
Pigment
production
;t++++t++4+4t4+ F441
Growth at
room temperature
Aerial
mycelium
Growth on
potato
+++ 4+ 44444
+++
_
Mycobacterium. In the case of the Strepto-
myces-designated cultures, 83 per cent pro-
duced sporulating aerial hyphae, 9 per cent
nonsporulating aerial hyphae, and 8 per cent
formed no aerial hyphae. The Nocardia-
designated cultures gave, with regard to
production of aerial hyphae, 24, 47, and 10
per cent, respectively. Among the 214 Nocar-
dia-designated cultures, 79 were recognized
as representing N. (Eppinger)
Blanchard. They all produced acid from glu-
cose and glycerol, and utilized acetate, mal-
ate, propionate, pyruvate, and succinate.
They all grew well at 28 and 35°C, and 88
per cent grew at 40°C. Eighty-six per cent
reduced nitrate to nitrite, 54 per cent hy-
drolyzed starch, and 34 per cent decomposed
gelatin. A large number of cultures desig-
nated as Nocardia (N. corallina, N. ery-
thropolis, N. globerula, N. lutea, N. opaca,
N. rhodni, N. rubra) were tentatively
signed by Gordon and Mihm to the myco-
bacteria under JW. rhodochrous (Overbeck)
asteroides
as-
nov. comb. (Table 2).
Of five species of aerobic actinomycetes
associated with various mycetomas, Mariat
(1957) recognized only N. asteroides and N.
brasiliensis as nocardiae; Streptomyces ma-
durae, S. pelletiert, and S. somaliensis were
considered streptomycetes, although
Mariat was not quite certain of their exact
systematic position.
Bojalil and Cerbon (1959) divided the
genus Vocardia into two different metabolic
groups: (1) Produces round colonies, adher-
ing to wall and bottom of tube; utilizes
gelatin as the only source of N and C, break-
ing it down into amino acids and giving an
alkaline reaction. NV. brasiliensis belongs to
this group. (2) Produces flaky growth easily
as
dispersed through medium; poor growth on
gelatin. N. asteroides belongs to this group.
A detailed examination of the variability
of different strains of two species of Nocar-
dia with regard to their ability to utilize
different
well as in certain other physiological and
‘carbon and nitrogen sources, as
biochemical properties, is reported in Table
2. Some comparative properties of several
nocardiae are given in Table 3.
Spalla (1958, 1959) eriticized the various
descriptions of Nocardia species on the basis
of an insufficient number of characters. He
suggested that the following properties be
used for characterization and classification
of nocardiae:
TABLE 4
Cultural and biochemical properties of certain species of Nocardia (Spalla, 1959)
Growth* on
dese auiseied
-Sse-asoonyyy
VSR [OIIIA[H
umtpaut
auIWIe-7-N
Other properties
uint,aoAUL
[euey
a0uR\sIsal ploy
Shae.
jo uoTVR[NSRO)
++t+4+ 01+
uljejas
JO SISA[OIPAPHL
++4++4+11+
yorrys
Jo SISA[OIPAP{
ayei}iu
jo_"uoTjONpay
Acid production from:
[Oq1oS-P
jouuopy
oat te oer ere abe atl
jouuuryy-p
++4+4+4
JOIII AT)
+++++4+4+4+
uypnuy
Pecilee ae oR lh ele st
asouyjey
asoyeyatL,
aso}IV'T
9s0}[P
asoiong
asoulqeiy-/
+++4+111
asouuem-p | | | | ++ I +
ssoumvyy | +++ | + |
S800 P |e ar ae oN Pl se
asoyElRet)-p
Organism
. asterotdes........
blackwellit .......
DUPURIYRIR Pl cen
Strain 959) 09224. 424
Strain 2p. soe oe
=
=
>
S
S
= Zi Sk
apricot; S = salmon.
«
c
rose; L = lilac; A =
colorless; R
yellow; O = orange; C
+ Now recognized as a Streptomyces.
Fy
THE GENUS
NOCARDIA 29
TABLE 5
Serological relations of major pathogenic actinomycetes (Gonzdlez Ochoa and Vasquez Hoyos, 1953)
Antigens
N. brasilien- 1
sis No. 447
VAD ODUSINIOR Od lteeace ne, shetssertan Sk Shy bo ee +
N. brasiliensis No. 447.............. +
N. brasiliensis No. 468 +
N. mexicana No. 414... ...........5.55: +
INES OSTETOUACSRINO:, NSir. oe.ctescye oes oi0mrs, 6 a oes +
INPASEENOUGES INO ANG). a ipttcaysc sauce cess +
Nie astern ovdesiNOe20) .f0n5ecsb es ss a a +
IN asteTvovdes NiOn O02b 6. 1355006050 +-00- +
N. asteroides No. 694................... +
INENGUPSOUEES NO. DOLL. oo. ek ete nates os +
INEACOnNCG NO G16). 2. 5. es ws oe eh +
N. leishmanii No. 1030............. +
DSEMITLA UT LCUN Oma Diet. 1. 2 5 he tna: sie ee cise ts
Se pelletventiNO., WISS.2 we... went ee _
SME PRUGANGUNO: GID. «ices icesicsue owes s =
S. somaliensis No. 1065............... —
SS; somalvensis No: 1064............-... =
ISIS OMMALLENSUSMINO. LOGOR 5. cad a+ cle See ee - —
S. paraguayensis No. 285.............. _
SPE OLOUISS NOM OU Siedanvaiee ss ur:.c sere hun ee ae —
GHUSC US NO OhOy tty: bee idem aes syle as -
im. wavenautae No: 9963... ......6.+:-:- —
’. asteroides S. madurae
Precipitin formation
S. pelletiert S. somaliensis S. paraguay-
No. 19 No. 412 No. 1185 No. 1065 ensts No. 285
a — — — —
aL == = = =
_ — — — ae
a = — — —
aa — — — a
a — — — =
4. =— — — —
+ — — — =
aa = — — —
a = = = =
+ — — = =
— — — =
— — — + —
== — — — —
= - - +
|
|
|
|
ap ct cea al
1. Color of growth on several synthetic and
at least one organic medium.
2. Acid production from various carbon
sources.
3. Heat resistance.
4. Size of terminal fragments.
5. Staining properties, notably acid-fast-
ness.
6. Diastase formation.
7. Gelatin liquefaction.
8. Reduction of nitrate to nitrite.
9. Coagulation and peptonization of milk.
10. Formation of aerial mycelium.
By utilizing these properties and consider-
ing the high degree of similarity, Spalla was
able to conclude that three mutants of NV.
rugosa belonged to the same species as the
parent, and that N. rugosa, N.
rubra, and N. blackwellii represent distinct
species, with a low degree of similarity.
asteroides, N.
A summary of some of the properties of
N. rugosa and three of its mutants, as well
as of certain other Nocardia species, is given
in Table 4. The marked difference between
the culture designated as NV. gardneri and the
other nocardiae may be noted, particularly
in utilization of carbon sources, starch hy-
drolysis, and formation of aerial mycelium.
These differences account for the fact that
N. gardneri is now recognized as a Strepto-
myces and not as a Nocardia.
Various serological reactions of different
species of Streptomyces and Nocardia are
given in Table 5. Further information on the
relation of Mycobacterium to Nocardia is
found in the work of Haag (1927) and
Gordon (1937).
Of the
Nocardia, the following three systems of
classification of species are presented here:
various treatments of the genus
30 THE ACTINOMYCETES, Vol. II
1. Classification of Nocardia* Species,
According to Jensen (1932a)
A. Partially acid-fast organisms with strongly re-
fractive cells; nonproteolytic, generally non-
diastatic; capable of utilizing paraffin.
I. Initial mycelium limited, rapidly dividing
into rods and cocci.
1. Slowly growing organism; cells 0.5 to
0.7 » in diameter.
Nocardia minima
2. Rapidly growing organisms; cells 1.0 to
1.2 win diameter.
a. Cystites not produced; rapid forma-
tion of cocci.
Nocardia corallina
b. Cystites produced; less rapid forma-
tion of cocci.
Nocardia salmonicolor
II. Initial mycelium well developed, richly
branching, dividing into rods and generally
into cocct.
1. Substrate growth soft, without macro-
scopically visible aerial mycelium.
a. Substrate growth red; may produce
variants with undivided substrate
mycelium and visible white aerial
mycelium, or yellow and white vari-
ants.
Nocardia polychromogenes
b. Substrate growth white to pale pink.
al. Growth in nutrient agar opaque,
cream-colored; coeci in broth
culture.
Nocardia opaca
b!. Growth on sugar-free nutrient
agar watery; no cocci in broth
culture.
Nocardia erythropolis
2. Substrate growth hard, yellow, with
white aerial sporophores
divide into chains of acid-fast cocci.
Nocardia paraffinae
B. Nonacid-fast organisms with weakly refractive
cells; no distinct formation of cocci. Diastatic.
I. Nonproteolytic; no aerial mycelium;
marked production of cystites.
Nocardia mesenterica
mycelium ;
Il. Proteolytic organisms.
1. Growth on nutrient agar with rapid
formation of unbranched diphtheroid-
like rods; no typical cystites; broth tur-
bid.
Nocardia actinomorpha
* Designated by Jensen as Proactinomyces.
bo
Growth with extensive mycelium on nu-
trient agar; simple unbranched rods not
formed; cystites present; broth clear.
Nocardia flavescens
2. Classification of Nocardia,* Accord-
ing to Krassilnikoy
A. Cultures colorless, some excreting a brown sub-
stance into the medium.
I. Aerial mycelium and spore-bearing hy-
phae produced in culture media.
1. Substrate and aerial mycelium ocea-
sionally forming septae; the hyphae
break up into long rods, 15 to 30 y;
spherical bodies not formed.
a. Saprophytes, found on dead sub-
strates.
Nocardia actinoides
b. Parasites, living in bodies of man
and animals.
Nocardia gedanensis
2. Mycelium producing frequent septae;
hyphae break up into short rods and
cocci.
a. Saprophytes.
al. Grow on protein media.
Nocardia actinomorpha
b!. Grow on paraffin.
Nocardia paraffinae
b. Parasites.
Nocardia bovis
Il. Cultures produce aerial sporophores on
the surface of the colonies, but no aerial
mycelium; sporophores are short, straight,
covering the surface of the colonies with
a thin, pale layer.
1. Cultures grow in organic media.
Nocardia albicans
~o)
2. Cultures grow in inorganic media.
Nocardia oligocarbophilus
III. Cultures not forming any sporophores or
any aerial mycelium; colonies smooth or
lichenoid.
1. Saprophytes found on dead substrate.
Nocardia albus
2. Parasites or symbionts living within
plants, animals, or man.
a. Organisms living in symbiosis with
plants, forming nodules on their
roots.
Nocardia alni
Nocardia myricae
Nocardia elaeagnii
* Generic name Proactinomyces used.
THE GENUS NOCARDIA 31
b. Organisms living in the bodies of
man and animals.
al. Anaerobes, living in absence of
oxygen.
Nocardia anaerobicus
b!. Aerobes, or microaerophilic
forms.
a2. Strict aerobes.
Nocardia lignierest
b?. Facultative aerobes.
a3’. Cells nonacid-fast.
Nocardia israeli*
b3. Cells acid-fast.
Nocardia muris
B. Cultures pigmented.
I. Cultures pigmented violet or blue, the
pigments diffusing into the substrate.
1. Well
produced, hyphae forming occasional
developed substrate mycelium
septae and breaking up into long rods,
20 to 30 w; colonies form a faint aerial
mycelium with straight sporophores;
spores cylindrical.
Nocardia gabritschewski
2. No aerial mycelium produced, hyphae
fre-
or substrate mycelium forming
quent and breaking up into
short rods and cocci.
septae
Nocardia cyaneus
Il. Red or orange pigment produced.
1. Mycelium forming occasional septae
and breaking up into long rods; some
give rise to a faint aerial mycelium and
short straight sporophores.
a. Saprophytes.
al. Cultures not forming any soluble
pigment in medium.
Nocardia fructifert
b!. Cultures producing a brown sub-
stance.
Nocardia polychromogenes
b. Parasites living in bodies of man and
animals.
Nocardia freerv
2. Hyphae forming frequent septae and
breaking up into short rods and cocci;
no aerial mycelium produced.
a. Saprophytes living on dead sub-
strates.
Nocardia ruber
b. Parasites living in bodies of man
and animals.
* See Chapter 2 for description of Actinomyces
israelit.
FiGuRE 11. N.
soil extract agar (Reproduced from: Gordon, R. E.
and Mihm, J. M. J. Bacteriol. 75: 240, 1958).
asteroides, strain 730, grown on
al. Cells acid-fast.
Nocardia asteroides
bl. Cells nonacid-fast.
Nocardia variabilis
III. Cultures citron-yellow or bright yellow.
1. Faint
sporophores
aerial mycelium with straight
and cylindrical spores
produced.
a. Saprophytes.
Nocardia flavescens
b. Parasites.
Nocardia somaliensis*
2. No aerial mycelium produced.
a. Saprophytes.
al. Cultures yellow or bright yellow.
Nocardia flavus
bt. Cultures citron-yellow.
Nocardia citreus
b. Parasites.
al. Cells acid-fast.
Nocardia farcinica
bl. Cells nonacid-fast.
Nocardia putoriae
IV. Cultures pigmented green.
1. Saprophytes.
Nocardia viridis
2. Parasites.
Noca rdia pyogenes
V. Cultures black.
1. Saprophytes.
Nocardia nige |
2. Parasites.
Now a rdia Sé ndate ns 1s
* Now recognized as a Streptomyces.
32 THE ACTINOMYCETES, Vol. II
3. Classification of Nocardia, According
to the system of Waksman and
Henrici*
A. Partially acid-fast organisms with strongly re-
fractive cells; nonproteolytic and generally
nondiastatic; capable of utilizing paraffin.
I. Initial mycelium fully developed, well
branching, dividing into rods and generally
into cocci.
1. Substrate growth soft, without macro-
scopically visible aerial mycelium.
a. Substrate mycelium yellow, orange,
or red.
al. Pathogenic.
a2. Substrate mycelium
buff, or pale yellow.
18. Nocardia farcinica
b2. Substrate mycelium yellow to
red.
6. Nocardia asteroides
b!. Not pathogenic.
a2, Paraffin decomposed.
42. Nocardia polychromo-
genes
b?. Cellulose decomposed.
13. Nocardia cellulans
b. Substrate mycelium white to pink.
al. Gelatin not liquefied.
a2. Growth on nutrient
opaque, cream-colored.
38. Nocardia opaca
b?. Growth on nutrient agar pink.
10. Nocardia calcarea
a3, Aerial mycelium on milk
white.
31. Nocardia leishmanit
b’. Pellicle on milk pink.
11. Nocardia caprae
e3, Pellicle on milk yellow.
9. Nocardia brasiliensis
white,
agar
d’. Causing galls on blueberry
plants.
53. Nocardia vaccinit
b!. Gelatin liquefied.
43. Nocardia pulmonalis
2. Substrate mycelium hard, yellow.
a. Aerial mycelium white; hyphae di-
vides into chains of acid-fast cocci.
40. Nocardia paraffinae
* This system was used, with certain minor
omissions and additions, in the last edition of
Bergey’s Manual.
b. Aerial mycelium not produced on or-
ganic media.
41. Nocardia petroleophila
Substrate growth cream colored, later
becoming yellow.
51. Nocardia serophila
Substrate growth hard, orange-yellow.
58. Nocardia variabilis
II. Initial mycelium very short, rapidly divid-
ing into rods and cocci.
a
Growth pink.
a. No cystites (swollen cells) formed.
al. No indigotin from indole.
16. Nocardia corallina
b!. Indigotin from indole.
26. Nocardia globerula
b. Cystites formed.
49. Nocardia salmonicolor
. Growth coral-red.
47. Nocardia rubropertincta
Growth white, tan, or pink.
a. No aerial mycelium.
a!. Growth tan.
15. Nocardia coeliaca
b!. Growth white.
28. Nocardia intracellularis
b. Aerial mycelium produced.
al. Growth frequently pinkish.
53. Nocardia transvalensis
b!. Growth never pink.
50. Nocardia sebivorans
Produces no pigment, no growth on
potato, coagulates milk.
19. Nocardia fastidiosa
. Nonacid-fast organisms with weakly refrac-
tive cells; no distinct formation of cocci; di-
astatic.
I. Nonproteolytic, although some give gela-
tin liquefaction.
i
Growth on agar pale cream.
a. Gelatin not liquefied; starch hy-
drolyzed.
35. Nocardia mesenterica
b. Gelatin liquefied; starch not hy-
drolyzed.
48. Nocardia rugosa
Growth on agar whitish.
4. Nocardia albicans
Growth on agar yellow.
20. Nocardia flava
Growth on agar green.
59. Nocardia viridis
Growth on agar yellow-green.
14. Nocardia citrea
6.
10.
14.
Il. Proteolytic,
. Growth
THE GENUS
Growth initially colorless, producing a
yellow-green pigment in 2 to 4 days.
54. Nocardia turbata
. Growth on agar dark brown and even
black.
a. No liquefaction of gelatin.
37. Nocardia nigra
b. Gelatin liquefied.
29. Nocardia tvorensis
. Growth consistency soft; aerial my-
celium sparse.
33. Nocardia lutea
consistency medium; aerial
mycelium profuse.
8. Nocardia blackwellit
Growth cream-colored to pink; aerial
spikes produced.
52. Nocardia sumatrae
. Growth grayish-yellow.
36. Nocardia muris
. Growth yellowish-orange.
55. Nocardia uniformis
Pigment on protein media deep brown.
44. Nocardia rangoonensis
Pigment on protein media light brown.
12. Nocardia caviae
although
some are only
weakly proteolytic.
Ie
Growth on nutrient agar with rapid
formation of unbranched diphtheroid-
like rods; no typical cystites; broth
turbid.
1. Nocardia actinomorpha
. Growth white, shiny or pale.
a. Dough-like consistency; breaks up
into short rods.
3. Nocardia alba
b. Membranous, myceloid growth.
32. Nocardia listert
Growth on nutrient agar with extensive
mycelium; simple unbranched rods not
formed; cystites present. Broth clear.
21. Nocardia flavescens
. Growth cream-colored.
a. Rapid liquefaction of gelatin.
al. No aerial mycelium.
25. Nocardia gibsonit
bl. Aerial mycelium scant, white.
56. Nocardia upcottit
b. Slow liquefaction of gelatin.
17. Nocardia dicksonii
. Growth rose-colored to bright red or
red-orange.
24. Nocardia fructifera
6. Growth pink to red.
NOCARDIA 33
FIGURE 12. N.
produced from: McClung, N. M. First Reg. Conf.
Asia and Oceania, Tokyo, 1956).
rubra, electron micrograph (Re-
a. Gelatin not liquefied.
2. Nocardia africana
b. Gelatin slowly liquefied or not at all.
46. Nocardia rubra
c. Rapid liquefaction of gelatin.
39. Nocardia panjae
7. Pigment on protein media light brown;
color of growth pink.
45. Nocardia rhodnii
8. Growth yellowish to golden brown.
22. Nocardia fordir
9. Growth yellow to reddish-brown; solu-
ble pigment brown to red.
30. Nocardia kuroishi
10. Growth tan to buff-colored.
23. Nocardia formica
11. Growth very limited on various media,
except potato.
27. Nocardia hortonensis
12. Occur in the sea; liquefy agar.
a. Growth yellow.
34. Nocardia marina
b. Growth yellow-orange.
7. Nocardia atlantica
13. Produce nodules on roots of plants.
5. Nocardia alni
In addition to the species included in the
and described below,
many more species of Nocardia have been
above classification
recorded in the literature, either under this
or under other generic names. Some are
listed in Chapter 13, under the incompletely
described forms. Others are synonyms. There
is no question that some of the Streptomyces
species described in Chapter 8 could just as
34 THE ACTINOMYCETES, Vol. II
well have been included among the Nocardia
forms. It is also possible some of those listed
as Nocardia could just as readily have been
included in the genus Streptomyces. Fre-
quently, the decision of the investigator as
to whether a certain culture should be in-
cluded in one genus or another was perfectly
arbitrary.
Descriptions of Nocardia Species*
1. Nocardia actinomorpha (Gray and
Thornton, 1928) Waksman and Henrici,
1948 (Gray, P. and Thornton, H. Centrl.
Bacteriol. Abt. II, 73: 88, 1928).
Morphology: Growth colorless, smooth,
consisting of long, branching filaments and
rods, 0.5 to 0.8 by up to 10 yw. In older cul-
tures, rods 2 to 3 uw long generally predomi-
nate. On some media, extensively branching
hyphae occur. Not acid-fast.
Nutrient agar: Round colonies, 1 mm in
diameter, convex, white, granular or resin-
ous; long arborescent processes from the
edge. No aerial mycelium.
Potato-glycerol agar:
wrinkled, pink to orange.
Egg medium: Growth raised, dry, smooth,
Growth dry,
salmon-buff.
Gelatin: Colonies round,
white, raised rim, edges burred. Liquefaction
saucer-like,
positive.
Nutrient broth: Turbid.
Milk: Coagulation and peptonization.
Starch: Hydrolyzed (diastase produced).
Sucrose: Inverted.
Nitrate reduction: Positive.
Phenol and naphthalene: Utilized.
Temperature: Optimum 25-30°C.
Source: Soil.
Remarks: Differs from N.
liquefaction of gelatin. No acid from glucose,
lactose, sucrose, or glycerol.
coeliaca in
2. Nocardia africana Pijper and Pullinger,
* For further details concerning some of the
species, the last edition of the Bergey Manual
should be consulted.
1927 “(Piper “As-andPullinger,-B. DD. 23:
Trop. Med. Hyg. 30: 153-156, 1927).
Synonym: Actinomyces africanus (Pijper
and Pullinger) Nannizzi Pollaceci, 1934.
Morphology: Substrate growth consists of
unicellular branching mycelium. Aerial my-
celium sparse, consisting of short, straight
hyphae. Not acid-fast.
Glucose agar: Colonies minute, red, dis-
crete, round and piled up into a pale pink
mass. Aerial mycelium thin, white.
Nutrient agar: Colonies discoid, flat, pink.
Glycerol agar: Growth made up of small,
heaped-up, colorless masses with pink tinge;
later, growth abundant, piled up, pale pink.
Potato agar: Growth bright red, made up
of small, round colonies with colorless sub-
merged margins, and piled up patches. Aerial
mycelium stiff, sparse, white.
“ge medium: Colonies small, colorless,
blister, partly confluent ; becoming wrinkled,
depressed into medium. Liquefaction shght.
Gelatin: Irregular pink flakes. No liquefac-
tion.
Milk: Surface growth bright red. Medium
gradually becomes opaque, reddish-purple,
with slow peptonization.
Source: A case of mycetoma in South
Africa.
3. Nocardia alba (Kxrassilnikov, 1941)
Waksman (Krassilnikov, N. A. Actinomyce-
tales. Izvest. Akad. Nauk. SSSR, Moskau,
1941, p. 1).
Morphology: Growth smooth or folded,
made up of white colonies of a dough-lke
consistency; shiny or pale. Substrate myce-
lium breaks up into short rods 2.7 by 0.7 to
0.8 uw, later changing into a mass of coccus-
like cells, 0.7 to 1 uw. Many cells are swollen,
others form side buds. Not acid-fast. No
aerial mycelium.
Synthetic agar:
sources of nitrogen; sugar, starch, or organic
Inorganic salts used as
acids utilized as sources of carbon.
Nutrient agar: Good growth. No aerial
mycelium.
THE GENUS
Gelatin: Growth good. Positive liquefac-
tion.
Milk: Coagulation and peptonization.
Starch: Rapid hydrolysis.
Cellulose: No growth.
Paraffin: No growth.
Nitrate reduction: Negative.
Sucrose: Inverted.
Source: Soil.
Remarks: Several subspecies were also
listed: N. chromogena, N. paulotropha (Ac-
tinobacillus paulotrophus Beijerinck, 1914),
N. alba lactica, N. diastatica, N. hoffmanni.
4. Nocardia albicans (Krassilnikov, 1941)
Waksman (Krassilnikov, N. A. Actinomyce-
tales. Izvest. Akad. Nauk, SSSR, Moskau,
1941).
Morphology: Growth red, hyphae break-
ing up into rod-shaped cells, 12 to 25 by 0.6
to 0.7 uw, up to 50 uw in length. Cells straight
or slightly curved, branching. Aerial myce-
lium not observed, except surface layer of
sporophores, which produce a velvety ap-
pearance. Multiplication by fission, seldom
by budding.
Nutrient
shiny.
Gelatin: No liquefaction.
Milk: No change.
Starch: Hydrolyzed.
Cellulose: No growth.
Nutrient broth: Growth poor; produces
agar: Growth good, smooth,
faint turbidity, which settles on bottom and
leaves a surface ring. No true mycelium.
Cells rod-shaped 5 to 10 uw, seldom 15 to 20 u.
Nitrate reduction: Negative.
Sucrose: Inverted.
Paraffin: Not utilized.
Source: Soil.
Remarks: Glycerol used as a
‘arbon, and nitrate as a source of nitrogen.
5. Nocardia alni (Peklo emend. v. Plotho,
1941) Waksman (von Plotho, O. Arch.
Mikrobiol. 12: 1-18, 1941).
Morphology: Mycelium contains fatty
NOCARDIA
source of
ey)
~
globules; cells filiform, branching, disinte-
Aerial
mycelium usually absent, but may be formed
grating into short rods and cocci.
on cultivation. Sporulating cultures form
white, spherical to oval spores.
Agar media: Substrate growth compact,
shiny, colorless or slightly brownish.
Gelatin: pellicle.
positive. Soluble pigment brownish.
Liquid media: Slimy surface film.
Tyrosine: Utilized as source of nitrogen;
color turns red-brown.
Cellulose: Not utilized.
Carbon sources: Produces lactic acid from
Surface Liquefaction
glucose and lactose.
Optimum reaction for growth: pH 6.0.
Habitat: Roots of the alder.
Remarks: Produces nodules on the roots
of the host plant. Said to bring about nitro-
gen fixation in symbiotic culture with the
plant.
6. Nocardia asteroides (Kppinger, 1891)
Blanchard, 1895 emend. Gordon and Mihm,
1959 (Eppinger, H. L. Beitr. Pathol. Anat.
9: 287, 1891; Blanchard, R. In Bouchard.
Traite Pathol. Gen. 2: 811, 1895; Gordon,
R. E. and Mihm, J. M. J. Gen. Microbiol.
20: 129, 1959).
Synonyms: Cladothrix asteroides Mppinger,
1890; Streptothrix eppingeri
1891; Actinomyces asteroides Gasperini, 1892;
Lossi- Doria,
Oospora asteroides Sauvageau and Radais,
1892; N. asteroides R. Blanchard, 1895. Ac-
cording to Ochoa and Sandoval (1956), N.
leishmanii. Chalmers and Christopherson,
and N. phenotolerans Werkam and Gammel
are synonyms of NV. asteroides. According to
Gordon and Mihm (1959), V. caprae (Silber-
schmidt) Waksman Henrici, N. ep-
pingert, N. minima, and N. sylvodorifera are
also synonyms.
Morphology :
growth, usually yellow to orange to orange-
red. Mycelium straight and fine; it breaks up
into small, coceoid forms and rods. Some
and
Typical actinomycete
strains are acid-fast ; others are only partially
36 THE ACTINOMYCETES, Vol. II
so. Aerial hyphae produced; they vary from
rudimentary to long branching. Some may
produce chains of spores (Fig. 11).
Sucrose nitrate agar: Growth thin, spread-
ing, orange. No aerial mycelium. No soluble
pigment.
Peptone-beef extract agar: Growth much
folded, light yellow, becoming deep yellow
to yellowish-red. No soluble pigment.
Yeast-glucose agar: Growth flat to
wrinkled, beige to dark pink. Some produce
white aerial hyphae.
Potato: Growth much wrinkled, whitish,
becoming yellow to almost brick-red.
Gelatin: Growth yellowish on surface. No
liquefaction.
Milk: Orange-colored ring. No coagula-
tion; no peptonization.
Starch agar: Growth restricted,
orange. No diastatic action.
Blood serum: No liquefaction.
Carbohydrate utilization: See Table 2.
Nitrate reduction: Positive.
Oxygen demand: Aerobic. According to
Chalmers and Christopherson (1916), it may
also grow anaerobically.
Temperature: Optimum = 37°C.
strains grow readily at 28°C.
Pathogenicity: Transmissible to rabbits
and guinea pigs, but not to mice.
Source: Human infections and soil.
Remarks: A number of strains of acid-fast
scant,
Some
actinomycetes isolated from human lesions
have deviated in certain particulars from
the description of N. asteroides, but not
sufficiently to warrant separation as different
species. According to Gordon and Mihm, all
strains of N. asteroides form whitish aerial
hyphae, these varying from rudimentary to
much branching. The following characteris-
tics were considered the most valuable in the
identification of the species: development of
filamentous colonies with aerial hyphae; fail-
ure to hydrolyze casein and to dissolve the
crystals of tyrosine and xanthine; acid pro-
duction from glucose and glycerol; failure to
form acid from arabinose, lactose, mannitol,
inositol, and xylose; utilization of acetate,
malate, propionate, pyruvate, and succinate,
but not benzoate.
Numerous varieties of this species have
been described. It is sufficient to mention
N. crateriformis, N. gypsoides, and N. pseudo-
carneus (Gordon and Mihm, 1957).
Type culture: IMRU* 3308; also 504.
7. Nocardia atlantica (Humm and Shep-
ard, 1946) Waksman (Humm, H. J. and
Shepard, K.S. Duke Univ. Marine Sta. Bull.
3: 78, 1946).
Synonym: Proactinomyces atlanticus.
Morphology: Hyphae long, branching,
breaking up into rods and cocci, 0.5 to 0.7 pu.
Involution forms in old cultures. Nonacid-
fast. Aerial mycelium not produced.
Synthetic and organic media: Growth
bright yellow or yellow-orange, smooth, com-
pact, of a doughy consistency. Colonies flat
with shghtly raised center. Soluble pigments
none. Mineral sources of nitrogen utilized.
Gelatin liquefaction: Positive.
Agar: Liquefied.
Milk: Coagulation rapid, acidified; pep-
tonization slow.
Nitrate reduction: Positive.
Starch: Hydrolyzed.
Cellulose: Decomposed.
Chitin: Decomposed.
Agar: Slowly digested.
Alginie acid: Decomposed.
Carbon sources: Acid produced from arab-
inose, xylose, rhamnose, raffinose, fructose,
galactose, gum arabic. No acid
from lactose, dulcitol, mannitol, or sorbitol.
Organic acids utilized: gluconie, lactic, malo-
not utilized: acetic,
sucrose,
nic. Organie acids
butyric, citric, ete.
Optimum temperature: 28-30°C.
Habitat: Marine algae and marine sedi-
ments.
* These designations represent the various cul-
ture collections where the type cultures are de-
posited.
THE GENUS
Remarks: Another closely related culture
has been described as Proactinomyces flavus
(see N. marina).
Nocardia blackwellii (Erikson, 1935)
Waksman and Henrici, 1948 (Erikson, D.
aa ee irch Council Spec. Rept. Ser. 203:
, 1935).
es
rod-like filaments,
sparsely branching hyphae. Aerial mycelium
short, straight; frequently large, round
ovoid cells are interposed in the irregularly
Growth consisting of short,
growing out into longer
segmented chains of cells.
Glycerol nitrate agar:
granular, irregular, thin, pinkish.
Growth extensive,
Nutrient agar: Growth confluent, wrin-
kled, with small, round, pinkish, discrete
colonies at margin.
Glucose nutrient agar: Growth abundant,
pale pink, in form of small conical colonies,
piled up, convoluted.
Potato agar: Colonies small, round, color-
less. Aerial mycelium white. Later, colonies
dull pink, submerged margins; few aerial
spikes, moderate aerial mycelium at top of
slant.
Gelatin: Colonies few,
along line of inoculation.
colorless colonies to 10 mm_ below
larger pink-yellow surface colonies
white aerial mycelium. No liquefaction.
Milk: Surface pellicle heavy, convoluted,
bright yellow. No coagulation; no peptoniza-
tion. Spalla states that milk is coagulated.
Carbon utilization: See Table 4
Source: Hock joint of foal.
Type culture: ATCC 6846;
colorless, minute,
Later, abundant,
surface;
with
NCTC 630
9. Nocardia brasiliensis (Lindenberg, 1909)
Castellani and Chalmers emend. Gordon and
Mihm, 1959 (Lindenberg, A. Arch. Parasitol.
13: 265-282, 1909; Castellani, A. and Chal-
mers, A. J. Manual of Tropical Medicine, 2d
Kd. William Wood & Co., 1913, p. 816;
Gordon, R. E. and Mihm, J.M. J. Gen.
Microbiol. 20: 129, 1959).
NOCARDIA 37
Synonyms: Gonzalez Ochoa (1945, 1953),
Gonzalez Ochoa and Sandoval ee and
Gordon and Mihm (1958) consider bra-
siliensis as the proper name for ae organ-
ism. <A. Boyd and Crutchfield,
N. pretoriana Pijper and Pullinger, f
mMextCanus
and
transvalensis Pijper and Pullinger, are con-
sidered as synonyms.
Morphology: Angularly
branched _ fila-
ments, bearing a few short straight aerial
hyphae; later, growth becomes spreading
and extensive. Aerial hyphae long and
branching to short and gnarled; divide to
form oval and cylindrical spores. Acid-fast-
ness variable, from 100 per cent to none.
Glycerol nitrate agar: Growth in form of
piled up pink mass. Aerial mycelium very
secant, white, at margin.
Glucose nutrient agar: Colonies pale buff,
umbilicated and piled up.
Yeast-glucose agar: Highly mutable.
Growth yellow to yellowish-orange, finely
wrinkled. Some strains produce no aerial
hyphae; other strains form mat of whitish
aerial hyphae. A few strains form amber to
brown soluble pigment.
Potato: Colonies small, raised, pale pink;
plug and liquid discolored. Later, growth
dull buff, dry and convoluted at base, round
and zonate at top of slant. Aerial mycelium
white.
Gelatin:
faction
Milk: Surface growth yellowish. Pale pink
growth up the wall of the tube. Solid coagu-
lum in | month; later, partly digested.
round,
raised pink
A few colorless flakes. No lque-
‘ge medium: Colonies few, color-
less in 3 days. Later, irregular,
mass, warted appearance; moderate degree of
liquefaction.
convoluted,
Serum Growth raised,
slightly pinkish.
agar:
Source: A case of mycetoma of the chest
wall in a South African native.
To guinea pigs and humans.
and
Pathogenicity:
Remarks: According to Gordon
38 THE ACTINOMYCETES, Vol. II
Mihm, NV. brasiliensis is distinguished from
N. asteroides by positive decomposition of
vasein and tyrosine and by acid formation
from inositol and mannitol. Additional char-
acteristics of this species include the follow-
ing: xanthine not decomposed; acid pro-
duced from glucose and glycerol; no acid
from arabinose, lactose, maltose, xylose, and
other sugars; utilizes acetate, citrate, malate,
propionate, pyruvate, and succinate, but not
benzoate.
According to Mariat (1958), N.
is characterized by a lack of proteolytic ac-
tivity; utilization of urea, (NH4)oSOs and
IXNO; as sources of nitrogen, not of nitrite.
Glucose, fructose, glycerol, and mannitol are
asteroides
utilized as sources of carbon, but not galac-
tose, xylose, maltose, and starch, although
paraffin is utilized.
N. brasiliensis is characterized by gelatin
hydrolysis; utilization of urea, (NH4)2SO,4
and KNO; as nitrogen sources; utilization
of glycerol, glucose, fructose, galactose, man-
nitol, xylose, and paraffin as carbon sources.
Type culture: IMRU 850.
10. Nocardia calcarea Metealfe and
Brown, 1957 (Metealfe, G. and Brown, M.
E. J. Gen. Microbiol. 17: 568-569, 1957).
Morphology: Gram-positive and partially
acid-fast. Mainly short rods (1.5 to 2.0 by
1.0 uw) together with unbranched aseptate
filaments up to 10 win length and occasional
branched filaments. filaments show
lemon-shaped swellings. After 4 days, short
rods snapping division typical of
corynebacteria; abundant unbranched fila-
ments (5.0 to8.0 by 1.0 w) and a few branched
filaments and cocci (1.0 uw) also present.
Many rods show differentiation of a swollen
Some
show
spore-like structure; these are usually formed
terminally or subterminally, one per rod. In
the filaments they are often formed in chains.
Colonies after 14 days consist of short rods,
cocel and rods with swollen cells.
Agar media: Colonies circular, raised, soft,
cream
without aerial mycelium; pink or
colored with distinct pink tinge when small.
Sucrose agar: Filaments are rare and short
rods and cocci are the predominant forms
throughout. Occasional lemon-shaped cells
are formed on all media.
Glucose and mannitol agars: Very long
branched filaments (10 to 25 w) present after
2 days, often with terminal chains of swollen
hyaline cells; these filaments usually frag-
ment, but a few persist.
Yeast extract-peptone agar: Cycle shorter
than on previous media, most of the fila-
ments having fragmented into rods and cocei
after 3 days.
Starch agar: Growth slight; no hydrolysis.
Milk: Heavy growth, turned alkaline; no
peptonization.
Gelatin: Beaded growth at top of stab.
No liquefaction.
Nitrate reduction: Positive.
Carbon utilization: Utilizes glucose, su-
crose, and maltose; poor growth with lactose.
Paraffin: Growth heavy in basal salts me-
dium with ammonium salt and flakes of
paraffin wax.
11. Nocardia caprae (Silberschmidt, 1899)
Waksman and Henrici, 1948 (Silberschmidt,
W. Ann. inst. Pasteur 13: 841-853, 1899).
Synonyms: This organism has been var-
iously described as S. caprae (Price-Jones,
1901). O. caprae (Sartory, 1923), A. caprae
(Nannizzi, 1934).
Morphology: Substrate growth forms thin,
branching filaments, breaking up into rods.
Aerial mycelium abundant on all media with
tendency to form coherent spikes; mycelium
very polymorphous, but
thicker segments appear. Slightly acid-fast.
not occasional
Brownish soluble substance.
Glucose-peptone-beef — extract
Growth irregular, bright pink, tending to be
agar :
heaped up. Later abundant masses, frosted
over with thin, white aerial mycelium.
Blood agar: Colonies minute, round, color-
less, aggregated in broad pink zones. Aerial
mycelium pale. No hemolysis.
Ege medium: Colonies tew, colorless, some
pink; aerial mycelium white. Later, growth
becoming dull pink, irregular, with scant
white aerial mycelium.
Potato: Growth abundant. Aerial myce-
lium pale pink. Growth becomes membra-
nous, considerably buckled.
Gelatin: Growth extensive, dull,
small raised patches of pink aerial mycelium;
later, ribbon-like, depressed. No liquefac-
with
tion.
Milk: Surface pellicle red. Solid coagulum;
no peptonization.
Starch: No hydrolysis.
Source: Lesions in goats.
Pathogenicity: To rabbits, guinea pigs,
and mice.
Remarks: According to Gordon and Mihm
(1959) N. caprae is a synonym of N. aste-
roides; according to Schneidau and Shaffer
(1957), however, the organism does not uti-
lize paraffin and does not hemolyze blood,
as shown on p. 68, Volume I.
Type culture: IMRU 783.
12. Nocardia caviae Snijders, 1924 (Snij-
ders, Geneesk. Tijdschr. Ned. Indie 64: 47,
75, 1924).
Morphology: Growth consists of initial
segmented hyphae, producing elements of
approximately even thickness, arranged in
angular apposition; later, forms long, pro-
fusely ramifying threads with strongly re-
fractile protoplasm. Aerial mycelium straight
and branching, the sporophores forming oc-
casional coiled tips, divided into eylindrical
spores.
Glucose agar: Growth piled up, convo-
luted, cream-colored to pale pink. Aerial
mycelium white.
Glycerol agar: Growth scanty.
Potato agar: Growth spreading, colorless.
Aerial mycelium dense white.
Egg medium: Growth heavily corrugated,
pale pink, with submerged margin. Aerial
mycelium dense white. After 3 weeks, color-
less transpired drops.
Potato: Colonies small, colorless. Aerial
mycelium white, powdery. Later, abundant,
THE GENUS NOCARDIA 3
raised, pale pink, confluent growth. Aerial
mycelium white. Plug discolored.
Gelatin: A few colorless flakes. No lique-
faction.
Milk: Surface growth colorless. Aerial
mycelium white. Coagulation positive.
Nutrient broth: Surface pellicle cream-
colored, wrinkled, extending up wall and
breaking easily; moderate bottom growth,
flaky. Medium discolored.
Source: Infected guinea pigs from Su-
matra.
Remarks: Schneidau and Shaffer (1957)
report that the organism is not acid-fast,
grows at 46°C, utilizes paraffin, liquefies
gelatin, hydrolyzes casein, liquefies starch,
and shows positive hemolysis.
13. Nocardia cellulans Metealfe and
Brown, 1957 (Metealfe, G. and Brown, M.
E. J. Gen. Microbiol. 17: 569-570, 1957).
Morphology: Gram-positive and partially
acid-fast. Branching aseptate filaments, 30
to 40 uw in length, often with swellings at in-
tervals; shorter filaments are less than 7.0 u
in length. Fragmentation commences about
the fourth day, the number of short rods
(1.56 to 2.0 by 1.0 uw) increasing rapidly.
Hyaline spore-like structures are produced
from the seventh day as slight terminal
swellings on the filaments. After 28 days, the
colonies consist of very short rods, cocci and
spore-like cells.
Agar media: Colonies raised, soft, without
aerial mycelium; cream-colored on most
media but characteristically bright yellow on
yeast extract-peptone agar.
agar: Filaments fragment
rapidly and are occasionally found after 28
days. Numerous Y-forms are found in older
less
Glucose
cultures.
Cellulose tubes: After 6 days there are
long (20 to 30 uw) branched and unbranched
with swellings.
filaments, terminal
Fragmentation is rapid and short rods and
many
cocci predominate during the stage of active
cellulose decomposition. Old cultures are
composed almost entirely of cocci.
40 THE ACTINOMYCETES, Vol. II
Milk: Acid and curd produced.
Gelatin: Beaded growth at top of stab.
No liquefaction.
Starch agar: Starch not hydrolyzed.
Nitrate reduction: Positive.
Carbon utilization: Glucose, sucrose and
maltose utilized; acid produced.
Paraffin: Growth heavy with trace of
yeast extract; no growth without yeast ex-
tract.
Type culture: ATCC 12,830.
14. Nocardia citrea (IKXrassilnikov, 1938)
Waksman and Henrici, 1948 (Krassilnikov,
N. A. Bull. Acad. Sci. USSR No. 1: 139,
1938).
Morphology: Growth yellow to yellow-
green, usually rough and folded, of a dough-
like consistency. No soluble pigment. In
young cultures, mycelium consists of very
very fine threads 0.3 to 0.5 mw in diameter.
After several days the cells break up into
short rods 0.5 by 1.5 to 5 w and into cocci
0.3 to 0.5 w in diameter. Cells are the small-
est of all the nocardias. Multiplies by fission
and bud formation. No aerial mycelium. Not
acid-fast.
Synthetic medium: Growth and pigmenta-
tion typical.
Nutrient agar: Growth good.
Gelatin: Liquefaction rapid.
Milk: Coagulation and peptonization.
Starch: Hydrolyzed rapidly.
Sucrose inversion: Positive.
Cellulose: No growth.
Mat: Weak growth.
Paraffin or wax: No growth.
Nitrate reduction: Positive.
Habitat: Soil and water.
15. Nocardia coeliaca (Gray and Thornton,
1928) Waksman and Henrici, 1948 (Gray,
P., and Thornton, H. Centr. Bakteriol.
Parasitenk. Abt. II, 73: 88, 1928).
Morphology: Growth in form of short,
curved, uneven-sided rods, 0.8 by 5 u; oc-
casional filaments up to 10 to 12 yp long; fre-
quently beaded, occasionally swollen or
branched; coecoid forms 0.8 to 1.2 w in
diameter are common, especially in older
cultures. Not acid-fast, or
shghtly acid-fast.
Nutrient agar: Colonies less than 1 mm
in diameter, round or irregular, raised, white,
resinous, edge irregular, burred. Deep col-
ones irregularly round or oval, edge slightly
broken. Slant filiform, convex, white, rugose,
resinous, edge undulate.
occasionally
Potato-glycerol agar: Growth dry, crum-
pled, orange-colored, becoming brown.
Gelatin: Surface colonies irregular, raised,
white, rugose, dull edge entire. Deep colonies
irregular, smooth or slightly broken. Stab
convoluted, buff-white to yellowish, dull.
Below surface the growth forms many ir-
regular hollow lobes, giving a glistening ap-
pearance, to a depth of 3 to 4 mm.
Milk: Shghtly alkaline after 5 to 7 days.
Nutrient broth: Turbid.
Nitrate reduction: None.
Phenol: Utilized.
‘ge medium: Growth
moist, verrucose, buff-colored.
Temperature: Optimum 22-25°C.
Source: Soil.
Remarks: No acid from glucose, lactose,
sucrose, or glycerol. No chromogenesis.
Hollow lobes produced in deep gelatin cul-
raised, smooth,
tures.
Type culture: ATCC 13181.
16. Nocardia corallina (Bergey et al., 1923)
Waksman and Henrici, 1948 (Hefferan, M.
Centr. Bakteriol. Parasitenk. Abt. II, 11:
459, 1904; Bergey et al., Manual, Ist ed.,
1923, p. 93).
Synonyms: Nocardia minima (Proactino-
myces minimus Jensen). Bacillus mycoides
corallinus Reader, 1926.
Morphology: Growth pink to
orange-yellow. Branching mycelium, gener-
ally curved. In older cultures, hyphae de-
generate generally into shorter rods and
red to
coeel. Not acid-fast.
THE GENU
Nutrient agar: Colonies smooth, pink,
shining; border lighter, edge filamentous or
with arborescent projections. As the colony
grows, the cells in the interior break up into
short rods and cocci which eventually form
the mass of the colony. Cells on the outside
remain filamentous, giving the colony a
burr-like appearance, and often forming long
arborescent processes. No soluble pigment.
Potato-glycerol agar: Growth filiform,
raised, dry, wrinkled, yellowish-brown to
coral red.
Gelatin: Surface colonies round, convex,
smooth, pink, shining, edge filamentous;
deep colonies, burrs. No liquefaction.
Milk: Reddish pellicle; milk becomes alka-
line.
Nutrient
scum.
Paraffin and phenol: Utilized.
Nitrate reduction: Positive.
Starch: Not decomposed.
Sucrose: Not inverted.
Kee medium: Filiform,
wrinkled, orange.
Temperature: Optimum 22-25°C.
Habitat: Soil.
Remarks: Some strains produce acid from
glycerol and glucose. No acid or gas from
broth: Usually turbid. Pink
raised, dry,
sucrose, maltose, or lactose. Phenol and m-
cresol are utilized. strains utilize
naphthalene. Krassilnikov (1949) reports for
his strains, good growth in high salt concen-
trations. Schneidau and Shaffer (1957) report
Some
positive acid-fastness, positive hemolysis
and urease formation.
1935)
Research
17. Nocardia dicksonii (Trikson,
Waksman (Erikson, D. Med.
Council Spec. Rept. Ser. 17: 203, 1935).
Morphology: Growth consists of long fila-
ments, sometimes wavy. Aerial mycelium
straight. Spores cylindrical.
Glycerol nitrate agar: Growth granular
and wrinkled, cream-colored. Medium deeply
discolored.
NOCARDIA 41
Glucose-asparagine agar: Growth wrin-
kled, colorless.
Potato agar: Growth abundant, colorless.
Ege medium: Growth yellowish-brown.
Starch agar: Strong hydrolysis.
Gelatin: Growth smooth, cream-colored on
surface. Liquefaction limited.
Milk: Coagulated, peptonized.
Habitat: Unknown.
18. Nocardia farcinica (Nocard, 1888)
Trevisan and De Toni, 1889 (Noeard, M.
E. Ann. inst. Pasteur, 2: 293, 1888: Trevi-
san, V., I. generi e le specie delle Batteri-
acee, Milan, 1889, p. 9).
Morphology: Growth yellow, of doughy
consistency. Markedly acid-fast.
Nutrient agar: Colonies yellowish-white,
irregular, refractive; mycelium filamentous.
Potato: Growth abundant, dull, crumpled,
whitish-yellow.
Gelatin: Colonies small, circular, trans-
parent, glistening. No liquefaction.
Milk: No coagulation; no peptonization.
Starch: No hydrolysis.
Nutrient broth: Clear, with granular sedi-
ment, often with gray pellicle.
Nitrate reduction: Negative.
Temperature: Optimum 37°C.
Pathogenicity :
domestic animals and guinea pigs.
Source: Cases of cattle farcy.
Type culture: IMRU 3318.
Pathogenic to certain
19. Nocardia fastidiosa Suter, 1951 (Suter,
L. S. Mycologia 43: 658-676, 1951).
Morphology: The organisms were similar
on all media studied, showing in general a
striking pleomorphism with coccoid, bacil-
lary, and filamentous forms. Many of these
were clubbed; some bore a striking resem-
blance to spermatozoa. Others were thick at
one end and tapered down to filamentous
tails. Filamentous forms were up to 25 uw in
length and, not considering clubs or swollen
portions, measured 0.2 to 1.2 u in diameter,
the average being about 0.8 to 1 uw. Stained
42 THE ACTINOMYCETES, Vol. II
preparations never showed a richly branch-
ing character, but an alternate type of
branching was fairly easy to demonstrate
after about 7 days’ incubation at 37°C. The
eoccoid forms were round, oval, or drop-
shaped. Neither septa nor nuclei were seen.
Spores were formed in short chains within
mycelial strands and were of the same diame-
ter as the mycelial strands. Similar spores
were also found singly and extracellularly.
The organisms taken from cultures were
partially acid-fast.
Growth on agar media: Colonies were slow
growing, appearing after 2 to 3 days as tiny
specks, which after 7 days’ incubation finally
achieved, but never exceeded, a size of about
1 mm in diameter. To the naked eye they
appeared = grayish-white, compact, and
smooth, and under low-power magnification
they appeared fluffy, raised, compact at the
center, and irregular and stringy at the edge,
due to the presence of radiating and tangled
filaments. Zigzag arrangements of elements
and branches, clubs, and curls were seen at
the periphery. The colonies were adherent
to the medium. The top surface was dry and
could be scraped off with a stiff wire loop,
but neither the whole colony nor any part
of it could be removed intact. On blood agar
after 7 days’ incubation, the colonies viewed
by transmitted light showed a characteristic
dense reddish center and a clear outer zone,
both areas being very sharply defined.
Optimum temperature: 37°C.
Oxygen requirements: The organism is a
facultative anaerobe, growing equally well in
the presence or absence of oxygen.
Proteolytic activity: The organism is non-
proteolytic. No odor of putrefaction was
perceived in any of the cultures; gelatin was
not liquefied; no growth occurred on serum
plates or on coagulated human serum.
Gelatin stab: No growth after 28 days at
17-20°C.
Potato: No growth at 37°C.
Carbon sources: Acid formed from glucose;
not from lactose, sucrose, maltose, or glye-
erol.
Nitrate reduction: Negative.
Habitat: Isolated from penile ulcer.
Remarks: N. fastidiosa is different from
previously described species of Nocardia in
the following ways: It is very fastidious in
its growth requirements. It does not grow
in synthetic media to which carbohydrates
have been added; it will not utilize paraffin;
it will not grow on potato or carrot; it will
not grow on acid-maltose agar nor on acid-
glucose agar; attempts to grow it on nutrient
agar and on BHI agar have given variable
results; its optimum temperature is 37°C. It
is delicate and is relatively slow growing; it
is never hardy or richly branching, and it
does not produce a surface scum or a con-
fluent or filiform growth. It is a facultative
anaerobe, differing in this respect from all
other Nocardias described with the excep-
tion of N. farcinica and N. rubropertincta.
The author distinctive
properties as follows: It produces a fairly
compact colony composed of tangled myce-
lium and exhibits radiating, clubbed,
branched, and curled elements at the per-
iphery. Fragmentation of the mycelium and
summarizes its
post-fission movement (zigzag arrangement )
occur at the periphery of the colony. Arthro-
spores are produced. Stained preparations re-
veal partial fragmentation into bacillary
and coccoid forms. Mycelial forms and spores
average slightly less than 1 mw in diameter.
Neither nuclei nor septa were observed.
Branching is of an alternate type. It is par-
tially acid-fast.
20. Nocardia flava (krassilnikov, 1988)
Waksman and Henrici, 1948 (Krassilnikey,
N. A. Bull. Acad. Sei. USSR No. 1: 139,
1938).
Not Proactinomyces flavus Humm_ and
Shepard.
Morphology: Cells at first filamentous,
0.7 to 0.8 » in diameter; later, they break
into long rods and then into cocei 0.7 w in
THE GENUS
diameter. Some strains form chlamydo-
spores. Numerous inflated cells of the bulbi-
form or fusiform type. Cell multiplication by
fission, cross wall formation, rarely by bud-
ding. Not acid-fast.
Synthetic agar: Colonies bright yellow or
golden.
Nutrient agar: Growth dirty, lustrous, or
rough and folded, of a dough-like consist-
ency, yellow to straw-colored. No soluble
pigment.
Gelatin: No liquefaction.
Milk: No coagulation and no peptoniza-
tion.
Starch hydrolysis: Shght.
Sucrose: Weak inversion.
Cellulose: No growth.
Paraffin and wax: No growth.
Fat: Weak growth,
Habitat: Soil.
21. Nocardia flavescens (Jensen, 1931)
Waksman and Henrici, 1948 (Jensen, H.
Proc. Linnean Soc. N. 8. Wales 56: 361,
1931).
Morphology: Substrate growth forms long,
branched, nonseptate hyphae, 0.4 to 0.6. u.
On nutrient agar and potato, septa are
formed, mycelium fragmenting, partly re-
sembling highly branched mycobacteria.
Aerial mycelium consists of fairly long
hyphae of the same thickness as the vegeta-
tive hyphae, not very much branched, with-
out spirals, often clinging together in wisps;
hyphae break up into fragments of variable
lengths, from 1.2 to 1.5 up to 10 to 13 ug,
showing an irregular, granulated staining.
Not acid-fast.
Nutrient agar:
and much wrinkled, first dirty cream-col-
Substrate growth raised
ored, later dark yellowish-gray, of a soft,
moist, curd-like consistency. Aerial myce-
lium absent. Soluble pigment absent.
Glucose agar: Substrate growth super-
ficial, wrinkled. honey-yellow, of a hard
and cartilaginous consistency. Aerial myce-
NOCARDIA 4:
—~
lium thin, smooth, white. Soluble pigment
yellow.
Potato: Substrate growth
and wrinkled,
raised
later
yellowish-brown, soft and smeary. No aerial
mycelium. No soluble pigment.
Gelatin: Liquefaction slow.
Milk: Coagulation;
with acid reaction.
Starch: Hydrolyzed.
Cellulose: No growth.
Paraffin: No growth.
Sucrose: Inverted.
Glucose broth: Rather secant growth.
Granulated, yellowish sediment; no surface
growth. Broth clear. No pigment. No acidity.
Nitrate: Sight or no reduction.
Source: Soil.
22. Nocardia fordi (Erikson, 1935) Waks-
man (Erikson, D. Med. Research Council
Spec. Rept. Ser. 203: 15, 1935).
Morphology: Substrate growth consists of
much
first cream-colored,
slow peptonization
filaments of medium length. Aerial myce-
lium straight, sparse. Small
spores on potato agar and starch agar.
Glycerol nitrate agar: Growth thin, ex-
short, oval
tensive, golden brown, convoluted.
Nutrient agar: small,
to golden, ring-shaped; later, heaped-up
Colonies creamy
patches, becoming golden brown and con-
voluted.
Ege medium: Colonies minute, cream-
colored, elevated, becoming golden brown,
raised.
Potato: Growth yellowish in thin terminal
portion, tending to be piled up. Aerial myce-
lium secant, white, at top of slant. Later,
growth abundant, golden brown, confluent,
partly honeycombed, partly piled up.
Gelatin: No visible growth, slight soften-
ing of gelatin; later partial liquefaction.
Milk: Surface ring brownish. Coagulation
positive.
Starch: Not hydrolyzed.
Source: Human spleen in a case of acho-
luric jaundice.
44 THE ACTINOMYCETES, Vol. II
23. Nocardia formica Harris and Wood-
ruff, 1953 (Harris, D. A. and Woodruff, H.
B. Antibiotics Ann. 1953-1954, 609-614).
Morphology: Mycehal development ex-
tensive, with no fragmentation of hyphae.
Ghost filaments and cytoplasmic condensa-
tions produced. In submerged culture,
straight and curved rods develop, exhibiting
the Y- and V-forms. Rods are 0.9 to 1.1 by
1.3 to 6.0 uw. Not acid-fast.
Sucrose nitrate agar: Growth very faint
or none at all.
Glucose-asparagine Growth fair.
Aerial mycelium grayish-white. Sporulation
poor.
Nutrient agar: Growth fair. Aerial myce-
lium none.
Peptone-glucose agar: Growth tannish-
colored. Aerial mycelium white to grayish,
gradually covering surface. Reverse side
dark brown. Soluble pigment brown.
Egg medium: Growth excellent, buff-col-
ored, convoluted, moist. No liquefaction.
Medium not discolored.
Starch agar: Starch hydrolyzed.
Gelatin: Liquefaction rapid. Soluble pig-
ment none. Growth settled on bottom of the
tube.
Potato: Very poor growth.
Nitrate reduction: Positive.
Casein: Hydrolyzed.
Paraffin: Not utilized.
Optimum temperature: 28°C; good growth
ats C:
Carbon utilization: No acid production in
agar:
organic media from glucose, glycerol, lac-
tose, maltose, and sucrose; acid produced in
media trom glucose, glycerol,
lactose, maltose, but not from sucrose.
inorganic
Antagonistic properties: Produces an anti-
biotic substance active against Trichomonas
and swine influenza virus.
Source: Isolated from an abandoned nest
of African ants in an imported mahogany
log.
24. Nocardia fructifera (krassilnikov,
1941) Waksman (Krassilnikov, N. A. Ac-
tinomycetales. Izvest. Akad. Nauk. SSSR,
Moskau, 1941).
Morphology: Growth not compact, mostly
of dough-like consistency, smooth or rough.
Hyphae breaking up into rods and in some
cultures into cocci. Not acid-fast. Aerial
mycelium well developed, whitish to rose-
colored. Sporophores long, straight or weakly
wavy, but not spiral-shaped. Spores cylin-
drical, 1.5 by 0.7 jx.
Synthetic agar: Growth rose-colored to
bright red. No soluble pigment.
Nutrient agar: Aerial mycelium weakly
developed or absent entirely.
Gelatin: Liquefaction slow.
Milk: Coagulation positive; peptoniza-
tion weak.
Sucrose: Inverted.
Starch: Hydrolysis weak.
Cellulose: Poor growth.
Paraffin: No growth.
Fats: Good growth.
Source: Soil.
Remarks: One strain was obtained as a
mutant of another Nocardia; another strain
was changed, after 8 months of cultivation,
into a typical Streptomyces. This species 1s
considered as a transition form between the
two genera.
25. Nocardia gibsoni (Erikson, 1935)
Waksman (Erikson, D. Med. Research Coun-
cil Spec. Rept. Ser. 203: 36, 1935).
Morphology: Young growing mycelium
branches profusely at short intervals, finally
grows out into long, frequently wavy fila-
ments. Property of producing aerial myce-
lium apparently lost.
Nutrient agar: Colonies small, cream-col-
ored, depressed, partly confluent, growing
into an extensive wrinkled surface layer.
Glucose nutrient agar: Growth cream-
colored, wrinkled, membranous.
Potato agar: Growth wrinkled, glistening,
membranous.
Blood agar: Colonies small, diserete, yel-
THE GENUS
lowish, irregularly wrinkled, clear hemolytic
zone.
small,
medium: Colonies round,
smooth, colorless, with conically elevated
1
Koo
55
centers.
Potato: No growth.
Gelatin: Dull white flakes sinking as me-
dium liquefies. Liquefaction rapid.
Milk: Coagulation positive; peptoniza-
tion limited.
Starch: Not hydrolyzed.
Source: Human spleen in a case of acholuric
jaundice. Injected into a monkey and reiso-
lated.
Type culture: ATCC 6852.
26. Nocardia globerula (Gray, 1928) Waks-
man and Henrici, 1948 (Gray, P. Proce. Roy.
Soe. (London) B 102: 265, 1928).
Morphology: Growth orange to orange-
buff. It consists of curved rods and _fila-
ments, | by 2 to9 uw, with many coccoid cells,
especially in old cultures. Rods and filaments
frequently irregularly swollen. Not acid-fast.
Capsules may be present.
Nutrient agar: Surface colonies irregu-
larly round, 3 to 5 mm in diameter, convex,
white, smooth, shining; edge undulate, erose.
Deep colonies, lens-shaped.
Gelatin: Surface colonies irregularly
round, | to 2 mm in diameter, convex, light
buff, smooth, shining. Stab: nailhead, irreg-
ularly round, convex, pinkish-white, smooth,
shining.
Potato-glycerol agar:
moist, smooth, pale pink.
Milk: Alkaline.
Nutrient and peptone broth: Turbid with
viscous suspension.
Nitrate reduction: None.
Growth filiform,
Ege medium: Growth spreading, raised,
moist, orange-colored.
Indole agar: Blue crystals of indigotin
formed.
Temperature: Optimum 25-28°C.
Phenol: Utilized.
Source: Soil.
NOCARDIA 45
Remarks: This organism resembles most
closely N. corallina. It is distinguished by
producing a more watery type of surface
growth, more nearly entire deep colonies,
and more particularly by the production of
indigotin from indole. No acid from glucose,
lactose, maltose, sucrose, or glycerol.
Type culture: ATCC 13,130.
27. Nocardia hortonensis (Erikson, 1935)
Waksman (Erikson, D. Med.
Council Spec. Rept. Ser. 203: 22, 1935).
Morphology: Substrate growth made up
of very slowly developing unicellular myce-
lium, composed of long slender straight
branching filaments. Aerial mycelium very
Research
sparse, forming straight hyphae only once
on potato. Not acid-fast.
Glycerol nitrate agar: Colonies coiled,
colorless, lustrous patches, isolated, with
central depression.
Nutrient agar: Growth very slow, as few
smooth, cream-colored, coiled colonies.
Glucose nutrient agar: Growth as coiled
and heaped up cream-colored translucent
masses.
Potato agar: Colonies colorless, blister;
later dull green heaped and coiled mass.
Medium becomes slightly discolored.
Potato: Colonies abundant,
umbilicated, round, some coiled in raised
colorless,
masses; later, liberal olive-green growth.
Aerial mycelium dense, velvety gray-green
at top of slant.
Gelatin: Colonies round, cream-colored on
surface and a few millimeters below. No
liquefaction.
Milk: Surface growth green; peptoniza-
tion positive. Color at first purple, later
brown.
Source: From pus containing typical
actinomycetic granules from parotid abscess.
28. Nocardia intracellularis Cutting and
MeCabe, 1949 (Cutting, J. T. and McCabe,
A. B. Am. J. Pathol. 25: 1-47, 1949).
Morphology: Filaments branched, becom-
ing fragmented, composed of bacillary ele-
AG THE ACTINOMYCETES, Vol. II
ments in series, 0.2 to 0.45 w in width. Liquid
cultures give branched colonies. The hyphae
do not form club-shaped tips, and lack
chlamydospores. Not discolored when
stained with fuchsin and treated with acid
alcohol.
Agar media: Colonies
wet-shining, smooth, and nonmucoid.
Potato agar: No growth.
Gelatin: Growth poor. No liquefaction.
Glycerol broth: White, mucoid masses
formed at bottom of tube. Medium remains
circular, raised,
clear.
Milk: Acid after 20 to 30 days.
Starch: Not changed.
Tyrosinase reaction: Absent.
Cellulose: Not decomposed.
Nitrate reduction: None.
Oxygen requirement: Does not develop
in the absence of oxygen, but grows in an
atmosphere having 10 per cent COs.
Paraffin: Used as the only source of car-
bon.
Temperature: Grows at 37.5°C and _ tol-
erates well temperatures up to 40°C.
Habitat: Observed in granuloma of in-
fected lymph nodes and in the feces of a
living patient whose death it ultimately
saused. Observed also at autopsy in widely
disseminated granulomatous lesions which
it produced.
Type culture: ATCC 13,209.
29. Nocardia ivorensis Combes, Kauff-
mann and Vazart, 1957 (Combes, R., Kauff-
mann, J., and Vazart, B. Compt. rend. 224:
821-824, 1582-1587, 1957).
Agar media: Substrate growth character-
ized by the black coloration of its coccoid
bodies, by their elongation, by their resist-
ance to dryness and to heat, by their cellulo-
lytic properties, and by their production of
an orange pigment on different media. Col-
omes at first whitish, centers becoming light
orange to brown, later black. Black circles
formed successively around the central circle
and finally becoming confluent. Later, sur-
face of colony is uniformly black, shiny, and
waxy; at the periphery, grayish, radiant
outgrowths develop in the agar, forming a
more or less regular fringe; on the surface of
this fringe brown, rapidly darkening, con-
centric zones appear.
Milk: Reddish-orange surface film, and
isolated colonies adhering to the walls of
the tube.
Potato: Growth orange,
places, with the appearance of coccoid forms.
Gelatin: Growth scant, slow; later the
culture is orange in color. Liquefaction
darkening in
positive.
Cellulose (filter paper or washed cotton),
moistened with the synthetic medium: Light
brown colonies appear in 2 days and turn
dark at 6 days, being entirely composed of
coccoid forms; later, colonies are entirely
black. Cellulose progressively disintegrates.
Paraffin: White colonies appear at 5 days.
They remain small and rapidly form coccoid
elements.
Nitrate reduction: Positive.
Remarks: Three isolated cultures differed
from each other mainly in the rapidity with
which they formed coccoid elements. Or-
ganism closely related to N. nzgra.
Habitat: Colony of
coast of Africa.
30. Nocardia kurotshi Uesaka, 1952 (Ue-
saka, I. J. Antibiotics (Japan) 5: 75-79,
1952).
Morphology: Aerial mycelium abundant.
Sporophores slightly curved at first, later
termites on ivory
turning around each other. Acid-fast.
Glycerol nitrate agar: Growth thin, pale
yellow. Aerial mycelium punctiform, white.
Soluble pigment yellow.
Nutrient agar: Colonies wrinkled, grayish-
yellow. No aerial mycelium. Soluble pigment
faint grayish-brown.
Glucose nutrient agar: Growth abundant,
at first yellowish-brown, then reddish-brown.
Aerial mycelium secant, white at margin of
THE GENUS
colonies. Soluble pigment red to wine-
colored.
Potato: Growth moderate, at first red or
brownish red, later dark brown. Aerial myce-
lium grayish white. Soluble pigment dark
brown.
Glucose broth: Red colonies forming
pellicle. Abundant, flocculent sediment.
Soluble pigment dark brown.
Gelatin: Growth yellowish-brown, sinking
into medium. No aerial mycelium. No lique-
faction. Soluble pigment yellowish-brown.
Milk: No coagulation. Slow peptonization.
Brown pigment.
Starch: Hydrolyzed.
Carbon source: Lactose well utilized.
Nitrate reduction: None.
Antagonistic — properties:
antibiotic neonocardin, active against vari-
Produces an
ous bacteria.
Source: Soil.
31. Nocardia letshmanii Chalmers and
Christopherson, 1916 (Birt, C. and Leish-
man, W. B. J. Hyg. 2: 120, 1902; Chalmers,
A. and Christopherson, I. Ann. Trop. Med.
Parasitol. 10: 255, 1916).
Morphology: Initial cells frequently swol-
len, large and irregular, aggregated in short
chains and then branching out into regular
narrow filaments; later entire colonies as-
teroid in appearance, very fine and close
angular branching, with aerial hyphae situ-
ated singly. Whitish-pink aerial mycelium
generally abundant with irregularly cylin-
drical conidia. Acid-fast.
Glucose nutrient agar: Colonies rounded,
elevated, red, with paler frosting of sparse
aerial mycelium. No soluble pigment.
Glycerol agar: Colonies small, round, pink,
tending to be umbilicated and piled up.
Aerial spikes stiff, white.
Potato agar: Colonies minute, colorless,
round. Aerial mycelium white, in patches.
Kgg medium: Growth colorless, confluent,
with little wart-like
studded projections
NOCARDIA 47
bearing stiff aerial spikes; later pinkish.
Aerial mycelium white. Medium discolored.
Gelatin: Colonies small, pink. No lique-
faction.
Milk: Surface growth; aerial mycelium
white turning pink. Coagulum solid, later
partly peptonized.
Pathogenicity: To rabbits, rats, and
guinea pigs.
Source: Fatal case of lung disease and
pericarditis in man.
Remarks: According to Gonzalez Ochoa
and Sandoval (1956), N.
synonym of .V. asteroides.
32. Nocardia listeri (Erikson, 1935) Waks-
mann (Erikson, D. Med. Research Council
Spec. Rept. Ser. 203: 23-24, 1935).
Morphology:
straight. Spores oval.
Glycerol nitrate agar: Growth abundant,
leishmanii is a
Sporophores short and
moist, cream-colored. Aerial mycelium pow-
dery, white, with exuded drops.
Calcium malate agar: Growth poor, in
form of a biscuit-colored membrane.
Nutrient Growth
cream-colored,
agar: smooth, moist,
margin depressed, center
elevated.
Glucose nutrient agar: Growth cream-
colored, glistening.
Potato agar: Growth extensive, colorless,
warted surface. Dirty pink coloration after
2 weeks. Scant white aerial mycelium after
4 months.
Potato: Growth abundant, dull, brownish,
wrinkled. Aerial mycelium white.
Gelatin: Surface colonies round, white;
after 45 days, confluent skin. Liquefaction
slight.
Blood agar: Colonies small, round, cream-
colored, with smooth, translucent surface.
No hemolysis.
Serum agar: Colonies small, irregular,
moist, cream-colored, tending to be heaped
up; later somewhat transparent.
Milk: Coagulated. No change in reaction.
Source: From human material.
48 THE ACTINOMYCETES, Vol. II
33. Nocardia lutea Christopherson and
Archibald, 1918 (Christopherson, J. B. and
Archibald, R. G. Lancet 2: 847, 1918).*
Morphology: Growth consists of irregular,
spreading, polymorphous colonies, compris-
ing swollen and segmented cells of all shapes
and sizes with markedly granular contents.
Later cells more monomorphous, the fila-
ments being arranged in angular apposition.
Glycerol nitrate agar: Growth in form of
yellowish-pink, wrinkled membrane.
Nutrient agar: Growth abundant, co-
herent, moist, pink, membranous with round
discrete colonies at margin.
Glucose nutrient agar:
reddish, smeary.
Potato agar: Small filamentous colonies
are formed; irregular angular branching.
Aerial hyphae few, isolated, short, straight.
Potato: Growth carrot-red, moist, thick,
granular in bands, partly raised and with
discrete round colomes. Aerial mycelium
sparse, colorless, very thin at top of slant.
Gelatin: Growth pale pink, wrinkled on
wall of tube. Colorless punctiform and stel-
late colonies in medium. No liquefaction.
Milk: Growth orange-red on surface and
at bottom.
Growth secant,
Ege medium: Growth poor, dull pink.
Source: Actinomycosis of the lachrymal
gland.
Remarks: According to Erikson, various
saprophytes, such as N. rubra and N. poly-
chromogenes, are closely related.
34. Nocardia marina (Krassilnikov, 1949)
Waksman (Humm, H. J. and Shepard, K.
S. Duke Univ. Marine Sta. Bull. 3: 76,
1946; Ixrassilnikov, N. A., Guide to the
identification of bacteria and actinomycetes,
Moscow, 1949).
Synonyms: Proactinomyces flavus Humm
and Shepard. Proactinomyces citreus marinae
Krassilnikov.
* Description after Erikson, D., Med. Res.
Council Spee. Rept. Ser., 203: 30, 1935.
Morphology: Growth smooth, — bright
yellow color, of a dough-like consistency.
Hyphae long, filiform, branching, breaking
down into short rods and cocci. No aerial
mycelium.
Synthetic and protein salt water media:
Good growth.
Gelatin liquefaction: Positive.
Agar: Liquefied.
Milk: No coagulation; peptonization posi-
tive.
Nitrate reduction: None.
Starch: Hydrolyzed.
Carbon sources: Acetic, lactic, and butyric
acids utilized. Acid formed from various
sugars.
Temperature: 25-30°C.
Habitat: Atlantic Ocean marine deposit.
35. Nocardia mesenterica (Orla-Jensen,
1919) Waksman and Henrici, 1948 (Orla-
Jensen, S. The lactic acid bacteria, 1919,
181; Jensen, H. L. Proc. Linnean Soc. N.S.
Wales 57: 373, 1932).
Morphology: Growth forms extensive my-
celium composed of richly branching hyphae
of a somewhat variable thickness, 0.4 to 0.8
u. No aerial mycelium. Later, hyphae divide
into fragments of varying sizes and shapes,
partly diphtheroid rods, but no real cocci.
There is, particularly in complex organic
media, a tendency to form large, swollen,
fusiform to almost spherical cells, up to 3.5
uw in diameter. These may stain intensely
with carbol fuchsin.
Glucose-asparagine agar: Growth fair,
raised, granular, very pale yellow, glistening.
Condensation water-clear.
Glucose-peptone agar: Growth excellent,
spreading. At first flat and smooth, pale
straw-yellow, perfectly hard and cartilagin-
ous, later raised and strongly folded, of a
loose, curd-like consistency, bright lemon-
vellow.
Potato: Growth
cream-colored smear.
Gelatin: Growth
seant, restricted, soft,
finely arborescent,
THE GENUS NOCARDIA 49
cream-colored in the stab. Surface colony
raised, folded, pale yellow. No liquefaction.
Milk: Small cream-colored granules along
the tube. No proteolytic action.
Starch: Hydrolyzed.
Cellulose: Not utilized.
Nutrient broth: Good growth; volumi-
nous, flaky, whitish sediment; broth clear.
Nitrate reduction: Negative.
Sucrose: Inverted.
Source: Fermented beets.
Remarks: Sodium _ nitrate,
phosphate, and asparagine are utilized, al-
though these are inferior to peptone as
ammonium
sources of nitrogen.
36. Nocardia muris (Schottmiiller, 1914)
de Mello and Pais, 1918 (de Mello and Pais.
Arq. Hig. Pat. Exot. 6: 183, 1918).
Synonyms: Streptothrix muris-ratti Schott-
miller, 1914. Streptobacillus moniliformis
Levaditi, 1925. Actinomyces muris Topley
and Wilson, 1946.
Krassilnikov, 1941.
Morphology: Slender branching filaments,
0.4 to 0.6 uw in diameter, breaking up into
rods and cocci. Often cells form long chains
of bead-like cells, with terminal club-like
Proactinomyces muris
swellings. Nonacid-fast.
Growth: None on ordinary media. Growth
occurs in presence of serum, ascitic fluid or
blood.
‘Nutrient agar: No growth.
Glucose agar: No growth.
Serum agar: Grayish-yellow, clear col-
onies, 0.2 to 0.3 mm in diameter, with
smooth, glistening surface and entire edge.
Yasily emulsifiable.
Gelatin: No growth.
Potato: No growth.
Milk: No effect.
Nitrate reduction: None.
Blood agar: Like serum agar. No he-
molysis.
Egg medium: Similar to growth on serum
agar. No liquefaction.
Oxygen: Grows aerobically. Grows also
under anaerobic conditions.
Acid production: Acid produced in serum
agar media, with glucose and salicin, some-
times with maltose and lactose.
Habitat: Parasite inhabiting nasopharynx
of rats. Isolated from body of patient bitten
by a rat.
temarks: Similar organisms by a variety
of names, such as A. putoriz, were also
listed. Above description based on data of
Topley and Wilson (1946).
37. Nocardia nigra (Krassilnikov, 1941)
Waksman (not N.
Chalmers; not Streptothrix nigra
Doria.) (Krassilnikov, N. A. Actinomy-
cetales. Izvest. Akad. Nauk. SSSR, Moskau,
1941).
Morphology: Growth rough, folded, shiny,
dough-like consistency. Cells thread-like,
breaking up readily into rods 2 to 10 by
0.7 w and cocci, 0.6 to 0.8 p. No aerial
mycelium. Gram-positive, not acid-fast.
nigra Castellani and
LOssl-
Agar media: Growth poor, at first color-
less or brownish, gradually becoming darker,
later dark brown and even black. Pigment
not excreted in medium. Many cells are
swollen to 3 uw in diameter.
Potato: Growth good.
Gelatin: No growth. No liquefaction.
Milk: No change.
Cellulose: No growth.
Paraffin and wax: No growth.
Nutrient broth: Small sediment produced.
Medium clear.
Carbon utilization: Utilizes glucose and
mannose, with formation of acid.
Source: Seldom found in soil.
Remarks: Culture rapidly loses its vi-
ability on continued cultivation.
38. Nocardia opaca (den Dooren de Jong,
1927) Waksman and Henrici, 1948 (den
Dooren de Jong, L. E. Centr. Bakteriol.
Parasitenk. Abt. II, 71: 216, 1927: Jensen,
H. L. Proc. Linnean Soc. N.S. Wales 57:
369, 1932).
oO THE ACTINOMYCETES, Vol. II
FIGURE 13.
N. opaca (N. erythropolis), showing scheme of branching;
glycerol nutrient agar, first
sketch 10 hours incubation; others at hourly intervals (Reproduced from: McClung, N. M. Lloydia 12:
153, 1949).
Svnonyms: Nocardia crystallophaga (Gray
and Thornton); N.
Thornton); Preactinomyces opacus (Jensen).
erythropolis (Gray and
Morphology: Growth lustrous, rose-col-
ored to red. Hyphae long, curved, irregular
and branching, breaking up into rods and
cocci. Not acid-fast. Gram-positive (lig. 13).
Potato-glycerol agar: Growth dry, rough,
crumpled, pink to buff-colored.
Gelatin: Colonies round, convex, whitish,
smooth, shining, with edges shghtly ar-
borescent. Stab: convex, whitish, smooth,
resinous, filiform, erose. No liquefaction.
ge medium: Growth spreading, smooth,
moist, salmon-colored.
Potato: Growth covered with tufts. of
aerial hyphae.
Milk: Grayish pellicle. No coagulation, no
peptonization. Reaction slightly alkaline.
Nitrate reduction: Positive.
Starch: Not hydrolyzed.
Sucrose inversion: Negative.
Carbon sources: Saturated, long chain
aliphatic hydrocarbons are utilized as sources
of energy.
Temperature: Optimum 30°C.
Source: Seldom found in soils.
Differs from WN.
N. polychromogenes in that the
Remarks: corallina and
cells are
much longer than those of the former and
THE GENU
much shorter than those of the latter. Erik-
son (1949) added the following character-
istics: Soft cream to pink growth on nutri-
ent agar media. On synthetic media, growth
colorless and thin, producing an initial myce-
lium, the hyphae dividing rapidly into short
rods; addition of 0.01 per cent MnSQO, stim-
ulated production of pale pink pigment.
Acid-fast cell elements predominated during
periods of maximum growth and free air
supply. A study of the morphology of NV.
opaca grown on hydrocarbons and _ fatty
acids has been made by Webley (1955).
39. Nocardia panjae (Erikson) Waksman
(Erikson, D. Med. Research Council Spec.
Rept. Ser. 203: 1935, 16-17).
Morphology: Substrate growth made up
of very small, round colonies; unicellular
mycelium with slender, branching filaments.
Aerial mycelium not visible on any medium,
but occasional isolated aerial branches.
Glycerol nitrate agar: Growth poor; scant
colorless patch.
Calcium malate agar: Growth colorless to
pink, spreading; later, bright red mass,
buekled and shining, colorless submerged
margin.
Nutrient agar: Growth irregularly piled
up, convoluted, colorless, easily detachable,
brownish.
Glucose agar: Small colorless coiled mass,
later heaped up as green growth.
Gelatin: Liquefaction rapid.
Milk: Surface growth pale green. Coagula-
tion and peptonization.
Potato agar: Growth as small elevated,
convoluted, colorless mass with purple tinge
in center.
Egg medium: Colonies small, round,
tough, colorless; margin well embedded.
Later, colonies elevated, warted, darkened,
medium discolored and broken. Slight de-
gree of liquefaction; medium dark brown.
Source: An uleer of the abdominal wall of
a patient in India.
)
NOCARDIA 51
1951)
H.
Wales 56: 362,
(Jensen,
1948 (Jensen,
Nexo:
40. Nocardia para ffinae
Waksman and Henrici,
Proce. Linnean Soc.
1931).
Morphology: Growth hard, firm, yellow-
ish, consisting initially of an extensive myce-
lium, with long, richly branching hyphae,
0.4 to 0.5 uw thick. After 5 to 6 days, numer-
ous end double
thickness, and divide into oval, spore-like
elements, 0.8 to 1.0 by 1.2 to 1.5 uw. Division
branches swell to about
starts at the tips of the swollen branches
and proceeds basipetally until most of the
hyphae appear divided. Primary septa have
not been seen in the hyphae. The spore-like
elements are markedly acid-fast. Aerial my-
celium white consisting of short, straight,
not very much branched hyphae, 0.4 to 0.6
u thick, which never show any differentiation
into spores (see also Erikson, 1949).
Sucrose nitrate agar: Growth very scant,
as thin colorless veil. Aerial mycelium trace,
white.
fair,
flat, growing into medium; pale ocher-yellow
Glucose-asparagine agar: Growth
to orange, with raised outgrowths on the
surface. Aerial mycelium secant, white.
Nutrient agar: Growth slow, somewhat
raised, ocher-yellow, hard, smeary surface
loose. Aerial mycelium scant, small white
tufts. No soluble pigment.
Potato: Growth mycelium granulated,
first pale yellow, later deep ocher-yellow to
orange. Aerial mycelium scant, white. No
soluble pigment.
Gelatin: No liquefaction.
Milk: No coagulation; no peptonization.
Starch: No hydrolysis.
Cellulose: Not decomposed.
Paraffin: Readily utilized.
Nitrate reduction: Negative.
Sucrose: Not inverted.
Liquid media (milk, broth, synthetic solu-
Small,
yellow to orange colors, firm but can be
tions): round granules of various
crushed into a homogeneous smear. In old
o2 THE ACTINOMYCETES, Vol. II
Ficure 14. Orskov’s motile Nocardia (Reproduced by courtesy of N. M. McClung).
broth cultures, a thick, hard,
brownish surface pellicle is formed.
Habitat: Soil.
41. Nocardia petroleophila Hirsch and
Engel, 1956 (Hirsch, P. and Engel, H. Ber.
deut. bot. Gesell. 69: 441-454, 1956).
Morphology: Grows slowly, but
orange to
abun-
dantly on all mineral media, faster in a
petroleum atmosphere. It grows on certain
organic media, but does not produce any
aerial mycelium. Substrate mycelium breaks
up readily into rods; ‘‘involution cells” are
formed abundantly. Mycelial threads are
long, monopodially branched, 0.6 to 1.2 u
in diameter, and contain ‘‘metacleromatic
granules,” readily stained with aqueous
methylene blue. Aerial hyphae have the
same diameter as the substrate hyphae,
little branched. Aerial mycelium wets with
difficulty. No aerial spores. Mycehal seg-
ments 1.2 to 5.0 by 1.2 to 12.5 uw. Gram-posi-
tive; not acid-fast.
Glucose-asparagine agar: Growth limited.
Colonies 0.3 mm, white, yellow reverse.
Aerial mycelium snow-white. Soluble pig-
ment none.
Caleium malate agar: Growth limited.
Nutrient agar: Growth limited. Colonies
whitish-yellow, 0.5 mm. No aerial mycelium.
Starch agar: Growth limited. Aerial my-
celum snow-white; reverse yellowish. Starch
not hydrolyzed.
Potato: No growth.
Gelatin: Growth in form of microscopic
colonies. No liquefaction. No pigmentation.
Milk:
No aerial mycelium. No coagulation; no pep-
Growth limited, yellowish, dry.
tonization.
Temperature: Optimum 25-28°C. Re-
sistant to drying.
Salt concentration: Resistant to high
concentration.
Habitat: Soil.
42. Nocardia polychromogenes (Vallée,
1903) Waksman and Henrici, 1948 (Vallée,
H. Ann. inst. Pasteur 17: 288-292, 1903;
Jensen, H. Proc. Linnean Soc. N.S. Wales
56: 79, 363, 1931).
Morphology: Growth bright red, coral-
red to red-pink, of a doughy consistency,
later becoming leathery. Aerial mycelium
whitish with pink hue. Substrate growth
THE GENUS
forms long wavy filaments, 0.4 to 0.5 by
70 to 100 uw, extensively branched but with-
out septa. Older cultures consist entirely of
rods, 4 to 10 uw, frequently in V-, Y-, or
smaller coceoid forms. Gram-positive, not
acid-fast, frequently showing bands and
granules.
Nutrient agar: Growth scant, orange-red.
No aerial mycelium. No soluble pigment.
Glucose agar: Growth raised, flat, glisten-
ing, rose-colored, later becoming folded and
coral-red.
Gelatin: Growth along stab thin, yellow-
ish, with thin radiating filaments. Surface
growth flat, wrinkled, red. No liquefaction.
Milk: Growth
colored surface granules, later forming a
thick, soft, orange-colored sediment. No
coagulation; no peptonization.
Starch: Hydrolyzed.
Paraffin: Utilized.
Cellulose: No growth.
Temperature: Optimum 22-25°C.
Source: Blood of a horse; soil in France
and Australia.
Remarks: Differs from NV. corallina in the
formation of very long filaments and in fili-
starts as small orange-
form growth in gelatin stabs.
Type culture: IMRU 3409.
43. Nocardia pulmonalis (Burnett, 1909)
Waksman and Henrici, 1948 (Burnett,
S. H. Ann. Rept. N. Y. State Vet. Coll.
1909-1910, 167).
Morphology: Mycelium acid-fast, espe-
cially in early stages of growth; breaks up
readily into oval-shaped cells. Growth
lemon-yellow with white aerial mycelium.
Consistency of colonies leathery.
Peptone-beef extract agar: Growth moist,
raised, in form of small, spherical colonies.
Glucose-peptone-beet extract agar:
Growth dull, whitish, convoluted.
Potato: Growth abundant, in form. of
small, translucent, round colonies, becoming
lemon-yellow. Later, growth becomes con-
NOCARDIA de
we
voluted or folded with chalky white aerial
mycelium. Color of plug brownish.
Gelatin: Colomes small, whitish, spheri-
cal; edges of colony becoming chalky white.
Limited liquefaction.
Milk: Colonies on surface of the medium.
Coagulation and gradual peptonization.
Pathogenicity: Nonpathogenic for rabbits
and guinea pigs.
Source: Lungs of a cow.
44. Nocardia rangoonensis (Erikson, 1935)
Waksman and Henrici, 1948 (Erikson, D.
Med. Research Council Spec.
203: 33-34, 1935).
Morphology: Growth consists of branch-
ing hyphae which segment and _ present
tept. Ser.
sipping and angular arrangement. Aerial
hyphae few, short, straight, later developing
into a profusely branching, long, waving
aerial mycelium. Not acid-fast.
Glycerol nitrate agar: Growth dull, mealy,
pink, wrinkled. Aerial mycelium
white. Medium slightly discolored.
Nutrient Colonies round,
umbilicated, raised, cream-colored to pale
secant,
agar: lobate,
pink. Later, colonies colorless, medium dis-
colored dark brown.
nutrient
voluted, coherent, cream-colored; medium
discolored. Later, growth wrinkled, biscuit-
colored, colorless margin. Aerial mycelium
on border, white. Soluble pigment dark
Glucose agar: Growth con-
brown.
Potato agar: Colonies small, round, lemon-
colored, partly confluent. Submerged growth
greenish. Aerial mycelium white. Medium
colored light brown.
Egg medium: Growth extensive, colorless.
Aerial mycelium in center, pale pink.
Gelatin: Colonies abundant, minute in
medium; larger, cream-colored colonies on
surface. Aerial mycelium white. Brown pig-
ment surrounding growth. No liquefaction.
Milk: Surface ring yellow. Coagulation
positive; peptonization partial. Soluble pig-
ment dark brown.
o4 THE ACTINOMYCETES, Vol. II
Source: Human pulmonary case of strepto-
thricosis.
45. Nocardia rhodnii (Erikson, 1985)
Waksman and Henrici, 1948 (Erikson, D.
Med. Research Council Spec. Rept. Ser.
203: 29, 1935).
Morphology: Substrate growth made up
of minute colonies, composed of hyphal seg-
ments arranged in angular apposition. Aerial
mycelium is short and straight. Later,
growth becomes extensive and spreading,
made up partly of long, branching filaments,
and partly of short segments exhibiting
branching, each giving rise to
aerial hyphae. Angular branching very
marked, delicate, spreading, herringbone
patterns being formed.
Sucrose nitrate agar: Colonies minute,
shipping
colorless, round.
Glucose-asparagine agar: Growth abun-
dant, coral-pink, convoluted, piled up.
Glycerol agar: Growth made up of dull
pink colonies, round and umbilicated, be-
coming piled up and deeper coral-red.
Potato agar: Growth abundant,
piled up and stiff. Aerial mycelium white at
pink,
top of slant.
medium: Membrane salmon-pink,
Kee
455
granular,
Gelatin: Colonies pale pink, in form of
surface pellicle and as sediment. Liquefac-
tion rapid.
Milk: Growth bright orange. Medium
unchanged.
Nutrient broth: Salmon-pink flakes im
sediment and colonies on surface. Medium
discolored.
Source: From reduviud bug, Rhodnius
prolixus.
Type culture: IMRU 653.
46. Nocardia rubra (Actinomyces ruber
sterilis Krassilnikov, 1949) Waksman (Kras-
siinikov, N. A. Guide to the identification of
bacteria and actinomycetes. Moskau, 1949).
Agar media: Growth red, smooth, nodular,
slightly lustrous with a gravel-like appear-
ance. No aerial mycelium produced under
laboratory conditions. Most strains form no
soluble pigment. Some produce a brownish
substance. Slightly acid-fast (Fig. 12).
Milk: Unchanged.
Krassilnikov examined 25 different strains
said to belong to this type, but differing
from one another in intensity of color and in
certain physiological properties. He believed
that under certain conditions of growth these
cultures would develop an aerial mycelium
and proper sporulation.
This group was divided by Krassilnikov
into four subgroups:
a. Flat, compact colonies, red to pink in
color, pigment insoluble. Gelatin not lique-
fied, milk unchanged or only peptonized;
starch not decomposed; nitrate not reduced.
b. Colonies raised, dry, crumbling at con-
tact with loop; red to brownish-red in color.
Ready growth on synthetic media. Gelatin
liquefied slowly; milk coagulated shghtly or
only peptonized; nitrate reduced to nitrite;
ready growth in paraffin and fats.
c. Colonies compact, growing deep into
substrate; pink to ight red in color; brown
substance excreted into substrate. Gelatin
shghtly liquefied; milk peptonized by some
strains, starch slightly decomposed; nitrates
not reduced to nitrites. Some cultures grow
slowly in cellulose. Do not grow on paraffin.
d. Flat or nodular colonies, growing com-
pactly into medium. Frequently develop
coremia on the surface; these consist of
thickly interwoven sterile hyphae. The cul-
tures grow poorly on artificial media. Gelatin
liquefied or only milk not
changed or only shghtly peptonized; starch
not slowly;
not decomposed. No growth on cellulose.
Ready growth on fats, paraffin, and wax.
temarks: IXrassilnikov this
eroup to comprise transition forms between
considers
Streptomyces and Nocardia. N. corallina 1s
believed to be a related form; a number of
synonyms are listed such as NV. agrestis, N.
THE GENUS
minima, and others. According to Schneidau
and Shaffer (1957), this form does not pro-
duce urease, whereas .V. corallina does.
47. Nocardia rubropertincta (Hefferan,
1904) Waksman and Henrici, 1948 (Grass-
berger, R. Miinch. med. Wochschr. 46: 343,
1899: Hefferan, M. Centr. Bakteriol. Parasi-
tenk. Abt. II, 11: 460, 1904).
Morphology: Growth in form of small rods
0.3 to 0.9 by 1.5 to 3.0 uw, showing angular
arrangement; later, nearly coccoid, 0.6 by
0.8 u. Tendency for branching on glycerol
agar, but branching does not occur com-
monly, though granules of aerial mycelium
are sometimes seen. Not acid-fast or varia-
ble.
Nutrient agar: Colonies small, granular,
becoming pink to red, depending on com-
position of agar.
Potato: Growth slow but excellent, inten-
sive red, becoming dull orange.
Gelatin: Colonies irregular with crenate
margin and folded surface, coral-red. Growth
in stab at first thin, then granular to arbores-
cent with chromogenesis. No liquefaction.
Milk: Surface seales thick, fragile, dull
coral-red ; Milk
somewhat viscid after 3 to 4 weeks.
Nutrient broth: Faint uniform turbidity
with salmon-pink pellicle, which is renewed
on surface as it settles to form a red sedi-
also sediment. becomes
ment.
Nitrate reduction: None.
Carbon sources: Utilizes benzene, petro-
leum, paraffin oil, and paraffin.
Temperature: Grows well between 20 and
Bw.
Oxygen requirement: Aerobic to faculta-
tively anaerobic.
Source: Isolated from butter, soil, and
contaminants of tuberculin flasks.
femarks: Mycobacterium-like —colomes
with coral to vermillion-red chromogenesis
on various media.
48. Nocardia rugosa DiMarco and Spalla,
.
~
NOCARDIA
1959 (DiMarco, A. and Spalla, C. Lab.
Ricerche Farmitalia, Milano, 1959).
Morphology: Hyphae short, 0.6 to 0.8 u
in diameter, wavy, later angular, radiating
from a center. After 20 to 24 hours, they
break up into rods 8 to 20 uw long. No
aerial mycelium. Nonacid-fast.
Glucose-asparagine agar: Growth color-
less, raised, moist, wrinkled.
Glycerol Abundant, — lichenoid
growth, dull cream. No consistency. Red-
dish-brown soluble pigment after 15 days.
Nutrient agar: Thick, cream-colored pel-
licle, rough and folded. Dough-like con-
agar:
sistency. After 15 days, brown soluble pig-
ment.
Potato agar: Growth smooth, folded, with
wrinkled and lichenoid portions. Colorless.
Soft consistency.
Milk: Coagulation; no peptonization.
Gelatin: Liquefaction positive.
Nitrate reduction: Negative.
Starch: Nondiastatic.
Sugar utilization: See Table 4.
Optimum temperature: 84°C.
femarks: Nonpathogenic. Produces vita-
min By».
Habitat: Cattle rumen.
Type culture: IMRU 3760.
This species was described further by
Spalla (1959) as follows: It produces a color-
less growth on glycerol, glucose, asparagine,
and N-Z-amine agars. Acid is produced from
qe
arabinose, glycerol, d-mannitol, and adoni-
glucose, galactose, ribose, rhamnose,
tol; but not from d-mannose, sucrose, malt-
inulin,
Ni-
trate is not reduced. Starch is not hydro-
ose, lactose, trehalose, rafhinose,
d-sorbitol, inositol, dulcitol, or salicin.
lyzed. Gelatin is liquefied. Milk is coagu-
lated. The organism will resist a temperature
of 60°C for 115 hours, but not 3 hours. It is
gram-positive. The terminal fragments are
1.56 + 0.266 u. No aerial mycelium is pro-
duced.
49. Nocardia salmonicolor (den Dooren de
18) THE ACTINOMYCETES, Vol. II
Ficure 15. N.
salmonicolor, growing on a hanging microdrop of liquid paraffin (a) surrounded by
sucrose nitrate salt solution; (b) same plus 1.5 per cent agar (Reproduced from: Webley, D. M. J. Gen.
Microbiol. 8: 71, 1953).
Jong, 1927) Waksman and Henrici, 1948
(den Dooren de Jong. Centr. Bakteriol.
Parasitenk. Abt. I, 7h: 216, 1927):
Morphology: Growth made up of short
mycelium disintegrating into rods and cocci.
Aerial mycelium sometimes stretching into
quite long filaments, with small refractive
granules. Many cells at the edge of the
colonies show club- or pear-shaped swellings,
up to 2.5 to 3.0 uw in width; many of these
swollen cells later germinate with the forma-
Acid-
fastness is found among the earlier stages of
tion of two more. slender sprouts.
growth, especially in some of the strains and
on some media (Fig 15).
Glucose-asparagine agar: Growth — re-
stricted, rather flat, edges lobate, surface
warty, glistening; at first pale orange, later
ocher-yellow; consistency crumbly.
Glucose-peptone-beet extract agar:
Growth excellent, of a doughy consistency,
spreading, flat, dense, edges lobate, surface
folded, glistening, yellow, gradually chang-
ing to salmon-pink and deep orange-red.
Potato: Growth good, raised, warty,
crumbly, glistening, at first buff, changing
to orange, and finally to almost blood-red.
Gelatin: Growth in stab secant, arbores-
cent. Surface colonies small, wrinkled,
orange. No liquefaction.
Milk: Pellicle of small cream-colored
granules, later a thick orange sediment. No
coagulation and no peptonization, although
milk appears slightly cleared, the reaction
becoming alkaline.
Starch: Not hydrolyzed.
Sucrose: Not inverted, although readily
utilized with sodium nitrate as a source of
nitrogen.
Paraffin: Readily utilized as a source of
carbon.
Cellulose: No growth.
Phenol: Not utilized.
temarks: A detailed study of the acid-
fast properties of this species has been made
by Erikson (1949). It closely resembles V.
corallina.
sebivorans Erikson, 1954
Pathol. Bacteriol. 68: 387-
50. Nocardia
(Erikson, 1D. J.
393, 1954).
Morphology : partially
Aerial mycelium white, with
characteristic nonwetting properties. Both
Gram-positive,
acid-fast.
substrate and aerial mycelium show spon-
THE GENUS
taneous segmentation into shorter and
longer cells of coccoid or bacillary dimen-
sions.
Agar media: Colonies firmly attached to
medium. No soluble pigment. No acid pro-
duced from glucose, mannitol, lactose,
sucrose, starch, raffinose, galactose, rham-
nose, sorbitol, maltose, dulcitol, glycogen, or
glycerol.
Sucrose nitrate agar: Growth fair; nitrate
utilized.
Gelatin: No liquefaction.
Carbon utilization: Paraffin is good source
of energy, also n-dodecane. Cresols not
utilized.
Temperature: Can withstand exposure to
90°C for 10 minutes in a phosphate buffer
suspension; denser suspensions withstood 3
minutes at 100°C (Erikson, 1955).
Habitat: Pus in a case of empyema.
Pathogenicity: Rabbits and guinea pigs,
slightly for mice.
Remarks: Cells have avidity for oily sub-
stances (lipophilic). Under unfavorable con-
ditions of growth, on the surface of solid
paraffin, large clubs and hexagonal cells are
produced (Erikson, 1955).
Type culture: NCTC 8595.
51. Nocardia serophila (Sartory — and
Bailly, 1947) emend. Waksman (Sartory, A.
and Bailly, C. Compt. rend. 224: 1533-1534,
1947).
Morphology: Hyphae
growth, much branched, occasionally curved,
0.4 to 0.5 uw in diameter. Nonmotile. Rarely
certain secondary branches are spiral shaped.
intracellular arthrospores.
produce angular
Terminal and
Gram-positive. Acid and alcohol resistant.
Growth characteristics: Grows with diffi-
culty on ordinary solid or liquid media;
grows well on serum or blood media.
Liquid peptone and serum media: Non-
viscous, cream-colored growth, detaches
from tube by agitation, medium remaining
clear.
Coagulated serum media: Growth in form
NOCARDIA 57
of small colonies, white at first, later be-
coming yellowish cream-colored.
Biochemical properties: Indole negative,
HS positive, neutral red not reduced. Sugars
like glucose, sucrose, lactose, galactose, and
mannose not attached.
Milk: Coagulation positive; peptonization
positive.
Coagulated serum: Not liquefied.
Nitrate reduction: Positive.
Oxygen demand: Strict aerobe.
Optimum temperature: 35-37°C.
Pathogenicity: Pathogenic to guinea pigs
and rabbits.
Habitat: Isolated from urine of patient
suspected of renal tuberculosis.
52. Nocardia sumatrae (Snijders emend.
Erikson) (N. cuniculi, Snijders, Geneesk.
Tijdsch. Ned. Indie 64: 47, 75, 1924).
Not Streptothrix cuniculi Schmorl, 1891;
not Nocardia cuniculi de Mello, 1919.
Morphology: Growth made up of large
swollen cells, giving rise to ramifying fila-
ments or to small chains of short, thick seg-
ments which branch out into more regular
hyphae. Sometimes the irregular elements
are beset with spiny processes before giving
rise to typical long, branching filaments.
Later the picture becomes more mono-
morphous, and short straight aerial hyphae
are borne, which presently exhibit irregular
segmentation.
Glycerol nitrate agar: Colonies small,
round, elevated, cream-colored, margins
depressed; becoming smooth, discrete, yel-
lowish.
Glucose-asparagine agar: Colonies minute,
colorless, becoming dull pink, partly con-
fluent and piled up. Aerial spikes few, stiff,
pink
Nutrient agar: Colonies small, round,
elevated, cream-colored, umbilicated and
radially wrinkled.
‘gg medium: Growth scant, pinkish,
smeary.
Potato: Growth coral-pink, dry, granular,
38 THE ACTINOMYCETES, Vol. II
covered to a considerable extent with white
aerial mycelium; piled up in center, discrete
colonies at margin.
Gelatin: Few flakes. No liquefaction.
Milk: Heavy yellow growth attached to
walls; solid coagulum in | month.
Nutrient broth: Surface colonies cream-
colored, scale-like; abundant, flocculent
bottom growth.
Source: Infected rabbits.
Remarks: Description given after Erikson
(1935)
53. Nocardia transvalensis Pijper and
Pullinger, 1927 (Pijper, A. and Pullinger,
B. D. J. Trop. Med. Hyg. 30: 153,(1927).
Morphology: Initial growth made up of
unicellular hyphae, the central branch being
frequently broader and showing’ dense
granular refractile contents. Aerial mycelium
white, forming straight hyphae, in some
cases becoming clustered into irregular
spikes. Colorless drops are exuded and a pink
coloration is produced in the densest part of
the growth on synthetic glycerol agar.
Angular branching with division of substrate
filaments. Aerial hyphae irregularly seg-
mented. Acid-fast.
Glycerol nitrate agar: Growth in form of
small, pink coiled masses. Aerial mycelium
thin, white.
Nutrient agar: No growth.
Glucose nutrient agar: Colonies raised,
granular, pink. Aerial mycelium white.
Potato: Growth dry, raised, convoluted,
pink. Aerial mycelium white.
Gelatin: Growth poor, in form of a few
irregular, colorless flakes. No liquefaction.
Milk: No change.
Starch: Not hydrolyzed.
vee
irregularly raised, coiled, dull pink mass.
medium: Growth in form of small,
Source: A case of mycetoma of the foot, in
South Africa.
Pathogenicity: To guinea pigs.
Remarks: According to Gonzalez Ochoa
and Sandoval (1956), N. transvalensis is ¢
synonym of NV. brasiliensis.
54. Nocardia turbata Erikson, 1954 (Erik-
son, D. J. Gen. Microbiol. 11: 198-208,
1954).
Morphology: Typical actinomycete, pro-
ducing a fine mycelium composed of slender
filaments, 0.1 « in diameter, which fragment
into rods and coccoid cells. Under appro-
priate conditions, many cells are motile.
Nonacid-fast.
Agar media: Growth good. Colonies small,
0.1 to 2.0 uw. Initially colorless, later pro-
ducing a yellow-green pigment on nutrient
agar. Pigment production favored by free
air supply, suppressed by acid reaction.
Broth cultures: Turbid when young; sedi-
mentation of cells later, when pellicle and
clarification of medium produced.
Acid production: Positive with glucose,
sucrose, maltose, lactose, galactose, xylose,
arabinose, glycerol, starch; negative with
mannitol, raffinose, rhamnose,
dulcitol (using a casein hydrolysate me-
dium).
Oxygen demand: Aerobic.
Optimum temperature: 20-30°C.
Nitrate: Utilized.
Gelatin: No hydrolysis, except in presence
of peptone (slowly).
Paraffin utilization: Negative.
sorbitol,
Habitat: Probably soil.
55. Nocardia uniformis Marton and
Szabo, 1959 (Marton, M. and Szabo, I.
Acta Microbiol. Acad. Sci. Hung. 6: 131-
135, 1959).
Morphology: The filaments of the sub-
strate mycelium rapidly break up into rods
and less frequently into coccoid bodies. The
size of these forms is 0.7 to 1.1 uw by 1.1 to
4.0 uw. In old cultures, swollen, club- or bottle-
shaped forms appear. The hyphae of the
aerial mycelium are
slightly developed
straight or waved, nonseptate, and contain
THE GENUS NOCARDIA 09
oval oidiospores. The mycelium is gram-
positive and is not acid-fast.
Agar media: The strains give nonbutyrous
colonies growing into the agar, with moder-
ately striated dull surface covered with
shghtly developed white powder-like aerial
mycelium. The color of the colonies is a
constant yellowish-orange; it turns
red or yellow; no soluble pigment is pro-
duced. In liquid synthetic media a surface
pellicle resembling agar colonies is formed.
Gelatin: No liquefaction.
Milk: No coagulation; no peptonization.
Sugar inversion: None.
Starch hydrolysis: None.
Nitrate reduction: Rapid.
Paraffin utilization: Shght or none.
Optimum temperature: 14-37°C.
Carbon utilization: Does not utilize man-
nose, dextrin, inulin.
Habitat: Deep layers B; horizon of saline
never
soils.
56. Nocardia wpcottit (Erikson, 1935)
Waksman (Gibson, A. G. J. Pathol. Bac-
teriol. 23: 357, 1920; Erikson, D. Med. Re-
search Council Spec. Rept. Ser. 203: 22-23,
1935).
Morphology: Growth forms long, straight
filaments, much interwoven and ramified.
Aerial mycelium slight, transient, slightly
acid-fast.
Glycerol Colonies small,
nitrate agar:
round, cream-colored, glistening; heavy
texture, margins submerged. Later, growth
very much convoluted and raised, broad
submerged margin; medium becomes slightly
reddish.
Calcium malate agar: Growth limited,
colorless, membranous, with undulating
margin.
Nutrient agar: Colonies smooth, shining,
round, cream-colored; margin submerged.
Aerial mycelium scant, white. Later, colonies
are large with greenish tinge; very sparse
aerial mycelium gradually disappears.
Glucose nutrient agar: Colonies smooth,
round, cream-colored; margin depressed,
centers elevated, hollow on reverse side;
later a coherent membranous growth, yellow-
ish.
Potato agar: Growth poor, in form of
small, blister Medium
shghtly discolored.
Egg medium: Colonies round, flat, color-
less, seale-like, some marked by concentric
rings and slightly hollowed in center. Growth
becomes yellow-brown.
Blood agar: Colonies large, drab, heavily
textured. No aerial mycelium. No hemolysis.
Gelatin: Growth abundant, flocculent,
cream-colored on surface. Gradual liquefac-
tion.
Source: From the spleen in a
acholuric jaundice.
colorless, colonies.
case of
57. Nocardia vacciniti Demarec and Smith,
1952 (Demarec, J. B. and Smith, N. R.
Phytopathology 42: 249-252, 1952).
Morphology: Growth in form of rods and
filaments, 0.4 u to 0.8 uw in diameter; granular
appearance when stained; eventually break-
ing up into bacillary forms. Few cells acid-
fast. Fat demonstrated by staining with
Sudan black B.
Sucrose nitrate agar: Growth scant, gray.
Nutrient agar: Growth poor, slow, granu-
lar, gray, sometimes pinkish in old cultures.
Gelatin: Growth dry, ribbon-like. No
liquefaction.
Starch nutrient agar: Growth dry, ribbon-
like, pinkish to orange. Hydrolysis of starch
positive.
Potato: Growth slow, spreading, raised,
oray.
Milk:
orange spots. No peptonization.
Nitrate reduction: Positive.
Carbon utilization: With ammonia as the
Growth dry, raised, gray with
source of nitrogen, acid formed from glucose,
sucrose, glycerol, and mannitol; reactions
variable with arabinose and xylose; no
growth on lactose or sorbitol.
Paraffin: Utilized.
60 THE ACTINOMYCETES, Vol. II
25-28°C;
scant at
Temperature: Growth best at
inhibited at 32°C; none or very
SAGE
Antibiotic activity: None.
Habitat: Causes formation of bud-prolifer-
ating galls on blueberry plants.
Type culture: ATCC 11,092.
58. Nocardia variabilis (Cohn, 1913) Waks-
man (Cohn, T. Centr. Bakteriol. Parasitenk.
Orig. 70: 290-306, 1913).
Morphology: Cells initially filamentous,
breaking up into rods and cocci. Nonacid-
fast.
Agar media: Colonies round, smooth and
lustrous, sometimes nodular; light brownish
in color to orange-yellow. Colonies attached
fast to the agar and partly removed with
some effort.
Gelatin: Growth orange-yellow. No lique-
faction.
Milk: Surface pellicle gradually becoming
light orange. No coagulation; no peptoniza-
tion.
Broth: Colorless surface pellicle, readily
dropping to bottom. Medium remains clear.
Temperature: Optimum 387°C; — good
erowth at 42°C; weak growth at 45°C.
Potato: Growth thin, colorless, becoming
in time yellow to orange-red; finally brown.
Blood media: No hemolysis.
Oxygen demand: Markedly aerobic.
Habitat: Isolated from bladder of cystitis
‘vases in man. Pathogenic to guinea pigs.
Remarks: Said to be similar to A. ochroleu-
cus, A. ochraceus, and A. carneus of Neukireh
(1902). According to Krassilnikov (1949), it
is closely related to NV. africana.
59. Nocardia viridis (Krassilnikoyv, 1938)
Waksman and Henrici, 1948 (Krassilnikov,
N: A. Bull. Acad. Sci. USSR2 Nomi: 139;
1938; Guide to the identification of bacteria
and actinomycetes. Moskau, 1949).
Morphology: Growth dark green in color.
Colonies of doughy consistency on certain
media (wort agar, potato), and compact on
others (nutrient). Pigment insoluble in
medium and in organic solvents. On protein
media, cells develop to form a thin mycelium
without visible walls. Cells often
branching, 0.7 to 0.8 uw in diameter, with
cross wall. After 5 to 7 days the cells break
up into rods 5 to 15 w long. Coeei not ob-
served. Cells multiply by fission, seldom by
budding. No aerial mycelium. Gram-posi-
tive. Not acid-fast.
Nutrient agar: Growth compact. Thin
mycelium produced.
Potato: Growth rough, much folded.
Gelatin liquefaction: Slow or none.
Milk: No coagulation; no peptonization;
some reports of positive peptonization.
Starch: Not hydrolyzed. Spalla (1939) re-
ported positive hydrolysis.
Sucrose: Not inverted.
Nitrate reduction: None.
Paraffin and fats: Growth good; less on
wax.
Cellulose: No growth.
Habitat: Soil.
Davis and Freer (1960) described as a new
species NV. salivae, an aerobic actinomycete
isolated regularly from the human mouth.
Strains of this species are characterized by
cross
their saccharolytic power, which thus distin-
guishes them from the typical soil nocardias
(see also von Magnus, 1947; Howell ef al.,
1959). Hirsch (1960) described N. saturnea,
an aerobic organism occurring in dust and
capable of utilizing petroleum.
Chapter 4
Characterization of Streptomyces Species
Important Characters to be Considered
for Recognition of Species and Va-
rieties of Streptomyces
In the identification and characterization
of Streptomyces species, the following char-
acters should be considered:
I
Morphological properties:
(a) Structure of substrate mycelium.
(b) Nature and formation of aerial my-
celium.
Structure and branching of sporo-
phores.
(d) Size and shape of spores.
(e) Surface of spores.
Cultural properties on various media:
(a) Growth characteristics.
(b) Development of aerial mycelium.
(c) Color of aerial and substrate my-
(¢)
celium.
Biochemical properties:
(a) Production of soluble pigments in
organic and in inorganic media.
(b) Utilization of carbon sources.
(ec) Starch hydrolysis.
(d) Sucrose inversion.
(e) Cellulose decomposition.
(f) Proteolytic activities: liquefaction of
gelatin, blood serum, and casein; co-
agulation and peptonization of milk.
Utilization of nitrogenous com-
pounds.
(h) Formation of oxidases: tyrosinase
and laccase.
(1) Reductases: nitrate reductase, sul-
fate reductase.
o> Or
ints
In characterizing
(}) Formation of antibiotics and vita-
mins.
(k) Formation of H»S in peptone-iron
agar.
Sensitivity to antibiotics:
(a) Sensitivity to pure antibiotic prep-
arations.
(b) Phenomena’ of “‘cross-resistance”’
and ‘‘cross-sensitivity”’ on artificial
media.
Sensitivity to phages.
Serological reactions.
Chemical composition.
Ecological properties.
Genetic relationships.
Age of culture. Information should be
submitted concerning the age of the cul-
ture when the particular properties were
studied and the manner in which the
culture has been kept in the laboratory.
Type cultures. The culture should be
deposited in a recognized collection and
the assigned number reported. Every
possible means for preservation of the
culture should be used. Preservation of
strains by lyophilization, soil culture,
mineral oil seals on active slants, or
storage in deep freeze is believed to re-
duce physiological changes to a mini-
mum. With the lyophilization technique
within the reach of even the small lab-
oratory, there is no excuse for an inves-
tigator, particularly one publishing on
designated strains, to ‘‘lose’’ his strains.
Streptomyces species,
only certain media should be used and well—
62 THE ACTINOMYCETES, Vol. II
defined conditions of growth recognized. Un-
necessary media and nonessential details had
better be left out to avoid cumbersome de-
scriptions and nonduplicable characteris-
tics that may apply to varieties or strains
rather than to species. A larger number of
media and more detailed descriptions may
not only fail to give additional information
but may complicate the description of the
species to such an extent as to render the
identification of freshly isolated cultures
difficult.
The composition of the media is usually
given first consideration for descriptive pur-
poses. According to Waksman (1958), Shi-
nobu (1958), and others, these media should
include: (a) at least three synthetic media,
preferably sucrose-sodium-nitrate-salt or su-
crose-ammonium-salt agar, glucose-.or glyc-
erol-asparagine agar, and calcium malate or
‘alecium citrate agar; (b) two or possibly
three organic media, such as nutrient. (pep-
tone-beef extract) agar, yeast extract-glucose
agar, potato-glycerol-glutamate agar, or oat-
meal agar; (c) three or four complex natural
media, notably potato plugs, gelatin, and
milk; (d) peptone-iron-yeast extract agar for
HS production; (e) tyrosine medium for the
tyrosinase reaction; and (f) a synthetic me-
dium for carbohydrate utilization.
Very few, if any, other media are required.
Liquid media, with the exception of those
previously listed, are better left out.
Morphological Properties
The method of study of the morphological
properties of the streptomycetes would in-
clude visual microscopic examination versus
electron microscopic studies; direct exami-
nation versus study of stained preparations;
and hanging drop versus agar surface cul-
tures.
STRUCTURE OF SUBSTRATE MYCELIUM
The substrate mycelium of a Streptomyces
does not, as a rule, segment spontaneously
into bacillary or coccoid forms. It produces
leathery or tough-textured growth, remain-
ing nonseptate and coherent even in old
cultures. Although no true septa are ob-
served in young cultures, it has recently been
reported that older cultures show at least
occasional septation. The compactness of
this substrate growth is responsible for the
fact that hquid media are always clear, un-
less the culture has been subject to phage
or lytic action.
NATURE AND
LIUM
PROPERTIES OF AERIAL MYCE-
The aerial mycelium is usually thicker
than the substrate mycelium. While the
morphology of the substrate mycelium is
usually undifferentiated, the aerial mycelium
of streptomycetes, under fixed conditions of
culture, shows sufficient differentiation that
a miscellaneous assortment of isolates can
be segregated into a number of groups hav-
ing like morphological characteristics. This
is one of the most important criteria for
classification in the genus Streptomyces. Sev-
eral aspects relating to the aerial mycelium
may be considered:
a. Gross macroscopic appearance. The rela-
tive abundance, structure (cottony, velvety,
powdery), formation of rings or concentric
zones, and pigmentation of the aerial my-
celium are important diagnostic criteria.
b. Microscopic properties. The microscopic
structure of the aerial mycelium gives a clear
picture of the morphology and reproductive
structures of the organism. The hyphae may
be long or short, with extensive or little
branching. The branching may be simple or
complex, monopodial or sympodial, broom-
shaped or verticillate. The fruiting bodies or
sporophores are short or long, occurring sin-
gly, in clusters, or as verticils; they are
straight, wavy, or spiral-forming. The spirals
or coils are either long and open or short
and compact. Spiral formation may take
place on one medium and not on others.
CHARACTERIZATION OF
|
i
STREPTOMYCES SPECIES 63
FiaureE 16. Schematic representation of different types of spirals produced by various Streptomyces
species; the spirals range from long to short, from compact to irregular (Reproduced from: Shinobu, R.
Mem. Osaka Univ. Lib. Arts and Ed. B. Nat. Sei. 7, 1958).
Before a culture is pronounced as forming
no spirals, therefore, it must be grown on a
variety of selective media that will allow
optimum sporulation. Drechsler (1919) sug-
gested use of the right-hand or left-hand
curvature of the spirals as a diagnostic fea-
ture, but
composition of the medium (Ettlinger ef al.,
1958: Shinobu, 1958). Verticil formation is
also an important characteristic of certain
this, too, is influenced by the
species; it can be simple or branching (pri-
mary or secondary verticils), the branches
being straight or forming spirals; but this
property as well is influenced to some extent
the medium. Al-
though nocardiae may produce sporulating
by the composition of
aerial filaments, these are never spiral-
shaped (lig. 16).
Waksman (1940, 1950) divided the or-
ganisms belonging to the genus Actinomyces
(largely the forms now included in the genus
Streptomyces) into the following five sub-
groups on the basis of the structure of the
sporulation hyphae.
I. Straight sporulating hyphae, monopo-
dial branching, never producing regular
spirals.
Il. Spore-bearing hyphae arranged in clus-
ters, or broom-shaped arising from
compression of the sporophores.
III. Spiral formation in aerial mycelium;
long, open spirals.
IV. Spiral formation in aerial mycelium;
short, compact spirals.
V. Spore-bearing hyphae arranged on my-
celium in whorls (verticils) or tufts.
(1941, 1949) the
genus Actinomyces (largely forms included
Krassilnikoy divided
in Streptomyces) on the basis of the following
properties: (1) spiral forming versus straight
sporophores; (2) alternate distribution of
sporophores on aerial mycelium versus verti-
)
cil formation; (8) spherical versus oval
spores; (4) colorless versus pigmented cul-
tures; (5) white versus colored aerial my-
celium; (6) saprophytes versus parasites.
Aiso et al. (1948) divided the genus Strep-
tomyces on the basis of the structure of the
aerial mycelium into six types:
I. Spirals not formed.
64 THE ACTINOMYCETES, Vol. II
F
Piate I. Diagrammatie representation of the morphology of the sporophores of Streptomyces (Re-
produced by special permission from Ettlinger et al. Arch. Mikrobiol. 31: 336, 1958).
a. Sporophores produce straight branching verticils on sterile aerial hyphae; S. retdculi type. b. Sporo-
phores produce open spirals as side branches on sterile aerial hyphae; S. purpurascens type. ¢. Sporo-
phores produce on sterile aerial hyphae verticils with open, more or less regular spirals; S. nowrsei type.
d. Sporophores formed as side branches on sterile aerial hyphae, straight or slightly wavy; S. phaeo-
chromogenes. e. Sporophores produced as verticils on sterile aerial hyphae, with open, irregular spirals;
S. echinatus. {. Sporophores monopodially branched, forming irregular open spirals at the end of long
hyphae; S. lavendulae. g. Sporophores monopodially branched, with open, regular spirals; S. parvullus.
CHARACTERIZATION OF STREPTOMYCES SPECIES 65
mM n fe} Pp
Prats II. Diagrammatic representation of the morphology of the sporophores of Streptomyces (Re-
produced by special permission from Ettlinger ef al. Arch. Mikrobiol. 31: 337, 1958).
h. Sporophores produce numerous short, monopodial branches on sterile hyphae; S. ramulosus. i. Sporo-
phores sympodially branched, forming tufts upon short main axes; S. griseus. k. Sporophores produced
upon a long, straight main axis, monopodially branched with frequent, regular spirals; S. fradiae. 1.
Sporophores monopodially branched, straight or slightly wavy; S. antibioticus. m. Sporophores mono-
podially branched along the whole axis with open, irregular spirals; S. erythraeus. n. Sporophores mono-
podially branched, with narrow, compact spirals; S. violaceoniger. 0. Sporophores sympodially branched,
in the form of trees with a long main axis; S. viridogenes. p. Sporophores monopodially branched, stiff
and straight; S. venezuelae.
69 THE ACTINOMYCETES, Vol. II
1. Straight, very little branching.
2. Wavy, abundant branching.
II. Spirals formed.
1. Spirals formed on the axis, irregu-
lar branching.
2. Spirals formed on branches in clus-
ters.
Verticils produced.
1. Verticillate branches entangled like
a net.
2. Verticillate branches formed on both
axis and branches, making primary
and secondary verticils.
Okami (1952) grouped the genus Strepto-
myces on the basis of formation of aerial
mycelium into the following types:
I. Spirals not formed.
1. Branches produced.
2. Branches not produced.
II. Spirals formed.
1. Spiral form mostly compact.
2. Spiral form mostly loose.
Shinobu (1958) eriticized the systems of
Aiso and Okami on the basis that insufficient
attention was paid to the nature of the me-
dium. In the system of Aiso et al. the distine-
tion between | and 2 of each type appeared
to him to be unclear, many forms belonging
to an intermediate type. Okami’s system
was considered as incomplete because the
1
formation of verticils was not taken into
consideration. Waksman’s system was_ be-
lieved to be comparatively better, but even
this system was criticized because cluster or
broom-shaped sporophore formation was not
considered as a sufficient characteristic, and
because a strain does not necessarily have
only one kind of spiral, but usually forms
various kinds of spirals which coexist (PI. IT).
Hesseltine et al. (1954) and Pridham et al.
(1958) considered the sporophore morphol-
ogy as reasonably stable under definite nu-
tritional requirements of the organisms.
Several morphological groups were sug-
gested. The components of each group were
considered as suggesting a logical natural
arrangement. The physiological data can be
’
used to produce ‘“‘species’”? or ‘‘species-
groups,” with morphology as a_ starting
point. Seven morphological sections were
created as subdivisions of the genus Strep-
tomyces. However, this system as well was
considered by Shinobu as having certain de-
fects because (a) culture media for morpho-
logical study were not examined thoroughly
enough; and (b) some of the sections may
often coexist In one strain.
Shinobu (1958) emphasized the following
morphological properties of the aerial my-
celium:
a. Outward appearance — of
(powdery, cottony, leathery).
b. Branching, especially the formation
and nature of verticils.
ce. Formation and nature of spirals.
d. Formation and shape of spores.
e. Thickness and length of mycelium.
Shinobu examined in detail the various
properties of the aerial mycelium, emphasiz-
ing again the need for suitable synthetic
media. He concluded that the nature of the
aerial mycelium is one of the most important
characteristics for taxonomic study, but that
it should be considered in connection with
composition of the medium. The aerial my-
celium was classified into the following three
mycelium
groups, from the standpoint of branching
and formation of spirals.
Group I. Straight or wavy aerial my-
celium, monopodial branching, never pro-
ducing spirals or clusters.
Group II. Spiral formation in the aerial
mycelium; long or short; loose or compact;
open or closed.
Group III. Verticil or cluster formation
in the aerial mycelium.
The loss of ability to form aerial mycelium
and sporogenous hyphae by certain Strepto-
myces cultures, on the one hand, and the
formation of aerial mycelium and_ sporo-
phores by certain species and strains of
Nocardia, on the other, led Bradley (1959)
to question the distinction between these
two genera. We have here simply another
CHARACTERIZATION OF STREPTOMYCES SPECIES 67
vase of natural overlapping between man-
made concepts of genera and species or the
improper labeling of cultures. Gordon and
Mihm (1957) emphasized that it is easy to
understand how a culture of NV.
that formed acid-fast coecobaecilli, rods, and
short filaments and growth
heavy, finely to coarsely wrinkled, cream-
colored to orange, and without noticeable
aerial hyphae, could be mistaken for a J yco-
bacterium. A culture of V. asteroides, how-
ever, that produced nonacid-fast, long, tan-
gled filaments and a cream-colored, pale
yellow, or beige growth thickly covered with
whitish aerial hyphae, could just as easily be
accepted as a Streptomyces.
Numerous other studies have been made
asteroides
whose was
of the micromorphology of the various spe-
cies and groups of Streptomyces, as in the
work of Burkholder et al. (1954), Hesseltine
et al. (1954), Ettlinger et al. (1958), Flaig
and Kutzner (1960), and others.
c. Spores. The spores, also called conidia,
produced from, or in, certain hyphae of the
aerial mycelium, or the ‘‘sporogenous hy-
phae,”” may be oblong, oval, or spherical.
Krassilnikov (1949) attached great impor-
ance to this character, as determined by the
light microscope, as a diagnostic feature.
Kriss et al. (1945) were the first to use the
electron microscope for study of spores of
Streptomyces. This was followed by the work
of Carvajal (1946); Kiister (1953); Flaig et
al. (1952, 1955, 1958); Baldacci and Grein
(1955); Grein (1955); Vernon (1955); and
others. Flaig et al. (1952) found that the
spores of some strains had smooth surfaces
while others had spiny surfaces. They later
detected spores with hairy and warty sur-
faces; the nature of the nitrogen source in-
fluenced the appearance of the spore surface,
organic nitrogen favoring spine formation.
Kiister (1955) classified Streptomyces spores
into two groups: (a) those producing a
smooth surface and (b) those having a rough
surface. Each of these groups was divided
into three subgroups, based on shape of the
spores. Thus there are spores with smooth
surfaces, with spines, with hairs, or with
warty protuberances, and spores that are
globose, long-ovoid, and cylindrical.
On the basis of a system of classification
that they had outlined, Baldacci and Grein
(1955) examined 50 strains of streptomycetes
with the electron microscope. Three types
of spores were recognized: (1) Oval, more or
less transparent spores; these were either
smooth or rough, the latter having a spiny
or hairy surface; the spines were either short
and thick or long and thin. (2) Round,
opaque spores, usually smooth. (3) Poly-
hedral spores, smooth and transparent, or
shghtly curved, wrinkled, and opaque. The
form of the spores was constant for the series
in Baldacci’s system. It can hardly be used,
however, as a species characteristic. A cor-
relation was observed (Pridham, 1959) be-
tween spore characteristics and sporophore
morphology (Table 6).
According to Preobrajenskaya et al. (1959,
1960), strains within one species as a rule
have a similar type of spore surface. Cultures
with a white, yellow, greenish-yellow, yellow-
gray, pink, or lilae mycelium have smooth
spores; those with a bluish aerial mycelium
have spiny and hairy spores, and species
with a gray aerial mycelium have spores of
all types. The diagnostic value of spore sur-
face characteristics was found to be dissimi-
lar for the various sections. The correlation
between the gray and bluish species and the
character of the surface of the spores was
considered as insignificant. Tresner ef al.
(1960) also emphasized the importance of
spore surface in classifying species of Strep-
tomyces; size and shape of spores of most
species were considered of limited usefulness
in taxonomic differentiation.
Lechevalier and Tikhonienko (1959) re-
ported that the spores of S. viridochromo-
genes were mostly elongated and those of S.
violaceus, spherical. The spines formed by
68 THE ACTINOMYCETES, Vol. II
TABLE 6
Morphology of sporophores and spores of streptomycetes* (Pridham, 1959)
eee AIREE Original strain no. Sporophore morphologyt BAN Ce
griseus B-1598 Carvajal SL 842 Straight to flexuous (RF) Smooth-walled
bikiniensis B-1049 Waksman 3515 Straight to flexuous (RE) Smooth-walled
CaNesCus 2419 Com. Sol. 811.0 Straight to flexuous (RF) Smooth-walled
venezuelac B-902 Gottheb 8-44 Straight to flexuous (RF) Smooth-walled
cinnamonen- _B-1588 Okami 154-T3 Hooks and open loops (RA) Smooth-walled
sis with many straight (RF)
sporophores
flaveolus B-1334 ATCC 3319 Hooks and open loops (aber- Spiny to hairy
rant) (RA)
albus B-1685 Waksman (ATCC Spirals (aberrant) (S) Smooth-walled
618)
hygroscopicus B-1865 NRRL isolate Spirals (8) Spiny
chartreusts 2287 Upjohn K-180 Spirals (8) Spiny
* Data taken from Carvajal, 1946; Vernon, 1955. All other data on spore morphology supplied by
K. L. Jones.
+ Sporophore morphology determined according to the methods of Pridham ef al., 1958.
Fiaurp 17. Coremia formation by certain Streptomyces species, X 500; stained by Corti’s method
(Courtesy of Dr. J. Giolitti, Milan, Italy).
CHARACTERIZATION OF STREPTOMYCES SPECIES 69
strains of both species differed cytologically.
The spines of the first seemed to be part of
the cell wall, whereas the spines of the sec-
ond seemed to be very superficial, appearing
only on the envelope. It was concluded that
spine formation is a stable characteristic of
the spores. The shape of the spores varied
with the composition of the medium. It was
suggested that complex organic media be
avoided for spore study.
COLONY STRUCTURE
The nature of the Streptomyces colony
growing on a standard agar plate has been
considered as among the important criteria
for characterizing and recognizing a particu-
lar organism. One may question, however,
the significance of this property in describing
a species. The morphology of the colony,
notably its general appearance, size, shape,
and texture, can all be readily determined
by superficial examination. Various other
properties may be recognized from a study
of the colony. Krainsky used the structure
of the colony, especially its size and shape,
as one of the major diagnostic criteria.
The superficial examination of gross col-
ony structure, particularly its texture, can
be of some help. Pridham* and others have
noted the following very general correlations:
1. Straight to flexuous cultures generally
are flat with a velvety, granular, or powdery
texture.
2. Loop cultures generally are flat with a
velvety to slightly granular texture.
3. Spiral cultures generally are elevated
with a somewhat floccose texture. Occasion-
ally, spiral cultures that are flat with a vel-
vety or granular texture may be seen. In
spiral cultures that are somewhat floccose
the sporulating aerial mycelium often con-
sists of long sterile hyphae with sporophores
branched oppositely, singly, or sometimes in
an apparent verticillate fashion.
* Personal communication.
FicureE 18. Sclerotium formation in Strepto-
myces (Prepared by H. Lechevalier, Institute of
Microbiology).
Figure 19. Sclerotium in species of Strepto-
myces, designated as new genus Chainia by Thir-
umalachar (Prepared by H. Lechevalier, Institute
of Microbiology).
4. Verticillate cultures generally are ele-
vated and floccose. Aberrant verticillate cul-
tures generally are flat with a velvety tex-
ture.
70 THE ACTINOMYCETES, Vol. II
Fiaure 20. Sclerotium in species of Strepto-
myces, designated as new genus Chainia (Prepared
by H. Lechevalier, Institute of Microbiology).
According to Krassilnikov (1955, 1959,
1960), the nature of the sporophore is a per-
manent property, being straight in the S.
globisporus group, spiral-shaped in the S.
coelicolor and S. violaceus groups. The same
constancy is true of the shape of the spores,
cylindrical versus oval or spherical, and of
the manner of spore formation, namely, frag-
mentation versus segmentation. Branching
of the sporophores, namely, vertical forma-
tion versus monopodial branching, 1s also a
constant, although a variable, property. No
single property, however, 1s sufficient to
characterize species. Coremia formation
(Fig. 17) is of no taxonomic significance;
however, production of sclerotia is believed
to be a constant property, in agreement with
Thirumalachar (1955), but not with Gattani
(1957), who denied its significance (Tigs.
18-20).
Cultural and Biochemical Characteristics
FORMATION OF PIGMENTS
Among the cultural properties of strepto-
mycetes, the color of the substrate growth
of the aerial mycelium and the spores and
the formation of soluble pigments in organic
and synthetic media play a major role in
characterizing species. This fact is amply
illustrated by the numerous specific epithets
referring to color that have been used to
designate various organisms. Unfortunately,
color characteristics vary greatly with age
of the culture, composition of the medium,
temperature of incubation, and nature of the
inoculum.
Before the introduction of synthetic me-
dia, it was a common practice to divide the
actinomycetes into two groups: (a) colorless
or nonchromogenic, and (b) pigment-produce-
ing or chromogenic forms. The latter com-
prised those organisms that produced deep
brown to black diffusible pigments when
grown on proteinaceous media. With the in-
troduction of synthetic media, 1t came to be
recognized that different organisms are able
to produce a great variety of pigments, rang-
ing from red to blue and from orange and
yellow to brown and black. Some are single
pigments, and others comprise two or more
constituent pigments. Some are water-sol-
uble and others are water-insoluble, as shown
in detail in Chapter 13 of Volume I. The
presence of oxygen is essential for pigment
formation. The pH of the medium greatly
affects the nature of the pigments, both in-
soluble and soluble.
When cultures are grown on optimum
sporulation media, the pigmentation of the
spores is highly significant; 1t may be ob-
served at an early growth stage, at maturity,
or only in old cultures, since changes in color
may occur with age of culture.
The formation of deep brown to black pig-
ments on organic media containing proteins
CHARACTERIZATION OF
and protein derivatives, notably the amino
acid tyrosine, 1s an important species charac-
teristic. Certain species may produce only
faint brown soluble pigments on organic
media, as well as on synthetic media. Differ-
ent cultures, especially on continued cultiva-
ion artificial media, will
variation in pigment production.
Since about one third of all species of
on show great
Streptomyces now recognized are melanin-
positive (Waksman, 1919; Skinner, 1938),
and since this property has been utilized ex-
tensively in the classification of actinomy-
cetes, a knowledge of this reaction is of great
importance. Gasperini (1891) first utilized
this property in dividing the aerobic Actino-
myces into A. chromogenus and A. albus. It
was later recognized, however, that melanin
production is characteristic of a large number
of species, including such important forms
as the plant pathogen S. scabies.
Beijerinck (1900, 1911, 1913) designated
as “melanin” the dark pigment produced by
A. chromogenus from peptone, although this
organism did not always produce the pig-
ment from tyrosine. He considered the pig-
ment as a catabolic product of the organic
nitrogen.
Lehmann and Sano (1908) first suggested
the expression “‘tyrosinase reaction.”’? They
used for their studies a tyrosine-containing
medium, melanin being known to be an oxi-
dation product of tyrosine. Waksman (1916,
1919,
that the production of a soluble dark pig-
ment on beef-peptone agar is due solely to
this reaction. Gelatin, containing no tyro-
1920) expressed considerable doubt
sine, gives the characteristic pigmentation.
Some species producing a typical dark pig-
ment on the beef-peptone agar may fail to
do so on synthetic media containing tyrosine.
Skinner (1938) recognized a difference be-
tween the dark pigment produced in peptone
media and not in tyrosine-containing syn-
thetic media and the black pigment pro-
STREPTOMYCES SPECIES rial
duced in tyrosine (or protein containing
tyrosine) media. The tyrosinase reaction
was, therefore, considered to be the proper
one
Shinobu (1958) attached great importance
to the “tyrosinase reaction”’ in the species
characterization of Streptomyces. Ettlinger
et al. (1958) also recognized the difference
between melanin formation and the tyrosi-
In line with the of
Beijerinck, Waksman, and Ettlinger et al.,
nase reaction. ideas
recognition will be given here to melanin
formation rather than to the tyrosinase re-
action.
The formation of yellow, red, blue, green,
and other soluble pigments is also highly
characteristic of the species growing on syn-
thetic media. There is considerable variation
in the intensity of these pigments, depending
upon the strain of organism. In view of the
fact that color standards are not always
available, Lindenbein (1952) suggested a
series of color designations which are simple
and convenient. This system in a modified
form is given in Appendix I.
Pigment formation is considered by Kras-
sinikovy (1960) as a constant specific prop-
erty, although the nature of the pigments
varies with the composition of the medium.
The color of the aerial mycelium is not con-
sidered as constant and is greatly influenced
by the composition of the medium (see also
Conn and Conn, 1941).
The variability in pigmentation of differ-
ent strains of S. aureofaciens was studied in
detail by Duggar et al. (1954) and Backus
et al. (1954), and is illustrated in Table 7.
UTILIZATION OF CARBON SOURCES
The ability of different species of actino-
mycetes to utilize as sources of carbon and
energy various organic substances, such as
ot
acids, fats, and amino compounds, can be of
carbohydrates, alcohols, salts organic
considerable diagnostic value. These studies
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CHARACTERIZATION OF
the early work of Waksman
(1919), who employed a synthetic solution
to which he added various carbon- or ni-
trogen-containing compounds. Liquid sub-
date from
strata were employed and cultures were
incubated under static conditions. The use
of static hquid substrata was later found to
give misleading results. In some cases, uni-
form inoculum distribution is not achieved
unless considerable care is taken. Numerous
studies (Pridham and Gottlieb, 1948; Bene-
dict et al.,
1955; ete.) indicate that solid
substrates and different basal media
later used (Table 8).
Hata et al. (1953) found a correlation be-
were
tween the groups and types of organisms
established on the basis of carbon utilization
and their antistreptomytic and antibae-
terial spectra.
Zahner and Ettlinger (1957) did not at-
tach major significance to the utilization of
carbon sources for characterizing species of
Streptomyces. They suggested that such in
formation be combined with other criteria.
None of the 125 cultures they studied could
use dulcitol, for example. The best carbon
sources for characterizing Streptomyces spe-
cies were found to be raffinose, /-xylose, d-
fructose, /-arabinose, and d-mannitol. Gor-
don and Mihm (1959) considered as a species
the
malate, propionate, pyruvate, and succinate.
characteristic utilization of acetate,
None of the actinomycetes produce gas.
Some are able to form acid, such as lactic,
from certain carbon sources. Gordon and
Smith (1954) used acid production from lae-
tose, maltose, xylose, and mannose as one
of the criteria for differentiating Nocardia
and Streptomyces species. Gordon and Mihm
(1959) later suggested for species separation
the use of acid formation from glycerol, glu-
cose, arabinose, erythritol, inositol, lactose,
maltose, mannitol, and certain other carbon
sources.
Carbon source utilization by 12 natural
STREPTOMYCES SPECIES
TABLE 8
variant strains of S. aureofaciens
(Backus ef al., 1954)
No apparent
utilization by
Utilization by
all variants
Utilization variable with strain
any variant Posi- | Nega-
tive tive
Sodium Glucose Galactose 11 i
acetate* Sucrose Sodium 10 2
Sorbose Maltose citrate*
Glycine Starches Levulose 9 3
Mannitol Dextrin Mannose 9 3
Arabinose Trehalose Lactose i) 3
Glycerol Magne- 7 5
| Sodium sium lac-
succl- tate*
nate* Xylose 6 6
Inulin
* Used at 0.4 per cent level, all others at 1 per
cent.
PROTEOLYTIC ACTIVITIES
Among the proteolytic activities of diag-
nostic value in separating genera, liquefac-
tion of gelatin, hydrolysis of casein, and pep-
tonization of milk are very important.
Species of Nocardia effect little, if any,
liquefaction of gelatin, whereas most species
of Streptomyces bring about liquefaction.
The rapidity of liquefaction varies greatly.
Some species show strong activity, and
others give only limited liquefaction. This
property, as well as milk peptonization, when
combined with the ability of the species to
produce brown to black pigments, provides
significant criteria for species characteriza-
tion.
In a study of 477 cultures of Streptomyces,
Stapp (1953) found only one that did not
liquefy gelatin. Detailed studies on proteo-
lytic activities of actinomycetes are found
in the work of Waksman (1919), Jensen
(1930), Gordon and Smith (1955), and Kutz-
ner (1956), as well as in Vol. 1, pp. 183-186.
Waksman (1919) reported that of 35 cultures
tested, 33 liquefied gelatin more or less rap-
74 THE ACTINOMYCETES, Vol. II
idly; when these tests were repeated three
times, considerable variation in the degree
of liquefaction was obtained. IXutzner (1956)
kept gelatin cultures for 31 days; only four of
210 failed to liquefy the gelatin. Reports of
inability to liquefy gelatin or coagulate or
peptonize milk of certain species may often
be questioned. Repeated tests with different
inocula might have shown different results.
Gordon and Smith (1955) suggested casein
hydrolysis as one criterion for the separation
of Streptomyces strains from those of Nocar-
dia.
Stapp (1953) reported further that in his
collection 18 cultures brought about coagula-
tion of milk without subsequent peptoniza-
tion, 431 caused coagulation and peptoniza-
tion, and 19 caused peptonization without
previous coagulation. A few cultures ocea-
sionally are found that cause no coagulation
or peptonization. One wonders whether re-
peated tests with different inocula might
show different results.
REDUCING PROPERTIES
The reduction of nitrate to nitrite has
been universally used among the criteria for
species differentiation. In view, however, of
the influence of nutritional factors upon this
reaction, and its quantitative rather than
qualitative nature, its significance in species
characterization may be questioned.
Proteolysis, starch hydrolysis, sucrose in-
version, cellulose utilization, and nitrate re-
duction were said (IXrassiinikov, 1960) to be
characteristic of all actinomycetes
and to have, therefore, no taxonomic signifi-
almost
cance. Sugar assimilation was considered,
however, as a more or less constant property.
UTILIZATION OF NITROGEN SOURCES
As a rule, utilization of nitrogenous com-
pounds has not been considered important
for species characterization. Shinobu (1958)
considered the utilization of urea, creatinine,
and certain amino acids as of some impor-
tance in species characterization. Gordon
and Mihm (1959) suggested that the ability
to attack casein, tyrosine, or xanthine can
be considered of some significance in charac-
terizing species.
The use of hydrogen sulfide production as
a taxonomic implementation in the differen-
tiation of Streptomyces species has been sug-
gested by Pridham (1948). Tresner and
Danga (1958) later modified the peptone-
iron agar medium. More than 900 strains
belonging to one or another of 94 species
were tested. There was a marked difference
in response from strain to strain within a
species; for example, 98 per cent of 227
strains of S. hygroscopicus were negative; 99
per cent of 112 isolates of S. lavendulae were
positive. When employed in conjunction
with other physiological, cultural, and mor-
phological criteria, hydrogen sulfide produc-
tion was said to give promise as an aid in
the systematics of the genus Streptomyces.
Sensitivity to Antibiotics
Actinomycetes, especially species of Strep-
tomyces, have been found in recent years to
produce a series of highly valuable chemical
substances, notably, antibiotics. This prop-
erty has come to be considered as highly
characteristic of a given species. The fact
that a large proportion of all the cultures of
Streptomyces isolated from natural
strates show some degree of inhibition of
growth of other microorganisms, when tested
sub-
on suitable media, suggested the ability to
form antibiotics to be of potential diagnostic
value. It is a question whether one is a
“Jumper” or a ‘splitter’? when one regards
the ability to produce an antibiotic as a
species rather than a strain characteristic.
Certain antagonistic strains belonging to the
S. griseus group, for example, are able to
produce various streptomycins and eyclo-
heximide. Others may form various actino-
mycins, grisein, streptocin, or candicidin.
It has been suggested that because the
CHARACTERIZATION OF STREPTOMYCES SPECIES
growth of homologous strains of an organism
is less inhibited than that of heterologous
forms, added weight could be given to the
potential diagnostic value of antibiotic pro-
duction. The application of the concept of
antibiosis as a major characteristic for the
speciation of actinomycetes is not generally
accepted, since the metabolism of these or-
ganisms is too complicated to give sharp
lines of autoinhibition. At most, it can be a
varietal rather than a species characteristic.
Krassilnikov (1950, 1958, 1960a, 1960b)
tended to overemphasize the importance of
antibiotics in species characterization of ac-
tinomycetes. He made two unjustified as-
sumptions: (a) every species synthesizes only
one particular antibiotic, (b) antibiotics do
not inhibit the growth of organisms belong-
ing to the species producing such antibiotics.
Many species and even individual cultures
are able to form a variety of different anti-
biotics; on the other hand, the same anti-
biotic may be produced by different organ-
isms. The growth of an organism may in
some instances actually be inhibited by its
own antibiotic, as with S. fradiae and neo-
mycin. Finally, a single culture may produce
mutants which either have lost the ability
to form a particular antibiotic or have gained
the ability to synthesize a totally different
antibiotic. It is somewhat dangerous to use
assumptions and generalizations as the basis
for species characterization. The importance
of considering antibiotic formation in the
systematization of actinomycetes has also
been emphasized by Gause (1955).
Actinophage Sensitivity
During the last 10 years several attempts
have been made to determine whether
“phage-typing” of actinomycetes might be
of some help in identifying unknown isolates.
The results obtained point to several facts
which must be kept in mind if one tries to
use this procedure for characterizing and
Classifying Streptomyces species.
~J
)
1. Actinophages vary greatly in their host
ranges.
a. Most which
tested against a large number of organisms
proved to be polyvalent; that is, they lyse
cultures that belong to different species or
even genera (different according to our pres-
ent species concept, which is based on the
system presented here). The data presented
by Bradley and Anderson (1958) might serve
as an illustration (Table 45, Volume I). The
activity of some phages upon members of
the genera Streptomyces and Nocardia led
these workers to question the validity of
separating these two genera, which have ac-
tually been placed in two different families
within the order Actinomycetales. Activity
of a polyvalent Streptomyces phage on two
Nocardia species was also found by Gilmour
et al. (1959). In a study carried out by St.
Clair and McCoy (1959), however, nine
phages which proved to be polyvalent
against several Streptomyces species failed to
attack any of the four Nocardia species
tested. The polyvalent character of other
phages tested against other species has been
shown also by other investigators (Hoehn,
1949; Chang, 1953, Rautenstein and Kofa-
nova, 1957; Gause et al., 1957; Mach, 1958;
Shirling, 1959a, b; Kutzner and Waksman,
actinophages were
1959a; Kutzner, 1960). Obviously, therefore,
a phage characterized by a wide host range
is usually of little value in species differ-
entiation, unless one is inclined to be a
“Jumper”? who demonstrates by the use of
a polyvalent phage that he is right and the
“splitters”’ are wrong.
b. Some phages have been found to be
specific, causing the lysis of strains of only
a few species or of only certain strains of one
species. In the latter case one might be justi-
fied in doubting the uniformity of the spe-
cies rather than in considering the phage as
“‘superspecific.”’ This seems to be definitely
true of the species ‘‘S. griseus,’’ various
76 THE ACTINOMYCETES, Vol. II
strains of which show a very different sensi-
tivity pattern against certain phages.
It is true, likewise, of the separation of
streptomycin-producing strains from grisein-
producers and other members of the former
S. griseus, which is now regarded as a species
group rather than a single species (Waks-
man, 1959). There have been various reports
concerning phages which are active upon
streptomycin-producing strains, but do not
attack grisein-producers or nonantibiotic-
forming cultures (Woodruff et al., 1947;
Waksman et al., 1947; Reilly et al., 1947;
Waksman and Harris, 1949; Hoehn, 1949;
Carvajal, 1953; Burkholder eé al., 1954).
Some streptomycin-producing cultures have
been found, however, that are resistant to
these specific phages (Okami, 1950; Carva-
jal, 1953; Kutzner, 1960). Other phages have
been discovered that are specific against S.
coelicolor (Kutzner and Waksman, 1959a;
Kutzner, 1960), S. lavendulae (Gause et al.,
1957; Shirling, 1959), and SS. olzvaceus (KXha-
vina and Rautenstein, 1958).
2. No general conclusion can be drawn
from the spectrum of a polyvalent phage in
regard to relationships between lysed strains.
However, a polyvalent phage can be useful
in taxonomic studies if it shows specificity
within a particular group of organisms that
are very similar in their other properties
and therefore hardly distinguishable. Fur-
ther, testing several polyvalent phages might
result in typical sensitivity patterns of the
organisms which might be of some value in
recognizing whether one has to do with
closely related or unrelated organisms.
3. Actinophages vary greatly when tested
against numerous strains. In some cases dif-
ferences in plaque counts might be due to
host range mutants which are present at a
concentration of 10° to 10° particles. These
mutants would attack a ‘‘new host”’ resist -
ant to the parent phage, as shown by Welsch
(1954, 1957) and Welsch et al. (1957). In
numerous other cases, however, the devel-
opment of host range mutants cannot ex-
plain the wide host range, and the phages
retain their polyvalent nature even after
several serial passages on heterologous hosts
(Chang, 1953; Shirling, 1959a; Bradley,
1959; Gilmour et al., 1959). It is necessary
to carry out phage tests with different dilu-
tions of the original phage preparation,
which should contain about 107 to 10° par-
ticles per milliliter.
4. A survey of the literature shows that
almost every investigator uses a different
medium for phage typing. The methods
comprise either spot tests or single plaque
counts. In a comparative study of different
media for phage typing, Kutzner (1960)
found that some phages gave similar plaque
counts on a variety of media. However,
counts of phages that formed tiny plaques
were found to be quite dependent on media
composition. Inorganic salt content of media
was found to influence plaque counts most
strikingly. Some phages gave no plaques on
media NaCl but gave high
plaque counts when plated on the same me-
dium without NaCl (with or without CaCl,),
while other phages showed higher activity
on NaCl than on CaCl, media. The expres-
sion of phage activity is apparently influ-
enced by a great many unknown factors.
One of the phages lysed some strains with a
medium containing NaCl and gave high
plaque counts, but showed no activity
against other strains on the same medium.
These results suggest that a medium found
containing
optimal for one host-phage system might be
quite unsuitable for another. Before a phage
is typed against a large number of strains,
an optimal medium must be developed. Bet-
ter still, tests should be carried out with
several different media, selected for their
usefulness with particular strains.
Serological Reactions
Use of serological techniques, particularly
those of agglutination and precipitation, has
CHARACTERIZATION OF STREPTOMYCES SPECIES Ub
been suggested for species identification of
actinomycetes. Aoki (1935-1936) was thus
able to differentiate between representatives
of three genera, Actinomyces, Nocardia, and
Streptomyces. By means of sonic vibrations,
Ludwig and Hutchinson (1949) prepared an-
tigen suspensions satisfactory for use in ag-
glutinin and precipitin reactions and for the
production of immune sera in rabbits. Use
of such suspensions in the identification of
actinomycetes was suggested by Yokoyama
and Hata (1953) and Hata et al. (1953). A
purified antigen of a streptomycin-producing
strain was found active against immune sera
of the same strain, but not against sera of
other antibiotic-producing organisms. These
investigators were thus able to establish the
close relationship of luteomycin- and chlor-
amphenicol-producing organisms.
Ochoa and Hoyos (1953) found a correla-
tion between microscopic morphology and
serological reactions which made it possible
to divide the actinomycetes into four
groups: Group 1, including species of Ac-
tinomyces and Nocardia; Group 2, made up
largely of Nocardia; Groups 3 and 4, com-
prising species of Streptomyces. Slack et al.
(1951), however, found that antisera pre-
pared with A. bovis brought about low titer
agglutination of Nocardia and of two species
of Streptomyces. They concluded that a close
antigenic relationship exists between mem-
bers of the genus Actinomyces and that there
is a group relationship among Actinomyces,
Nocardia, and Streptomyces. Okami (1956)
found definite antigenic relationships be-
tween strains of closely related forms of S.
lavendulae. See also Tanaka et al., 1959.
Chemical Composition
A detailed study of the chemical composi-
tion of cells of actinomycetes has been pre-
sented in Volume I (pp. 158-163).
The occurrence of specific chemical com-
pounds in the cells of the organisms suggests
possible differentiation between groups of
actinomycetes. This is true, for example, of
the occurrence of diaminopimelic acid, a
constituent that may prove to be of generic
rather than specific significance. Romano
and Sohler (1956) and Sohler et al. (1958)
have shown that cell walls of streptomycetes
can be solubilized by lysozyme, suggesting
the presence of a mucopolysaccharide; on
the other hand, cell walls of nocardiae do
not possess this property.
Ecology
The natural substrate of an organism, es-
pecially diseased plants or animals, and com-
posts of stable manures and plant residues
at high temperatures, is of some systematic
significance. Various attempts have been
made to utilize the ecological characteristics
of the actinomycetes as a basis of classifica-
tion. Thus, the following rather broadly de-
fined ecological categories have been pro-
posed at various times to classify actino-
mycetes:
a. Animal parasites.
b. Plant parasites.
c. Soil inhabitants.
d. Water inhabitants.
e. Mesophilic forms.
Thermophilic forms.
Inhabitants of acidic (pH 3 to 6.5) sub-
strates.
h. Inhabitants of neutral to alkaline
strates (pH 6.5 and above).
The temperature at which an organism is
oq
sub-
erown greatly affects the nature and amount
of growth, the nature and extent of sporula-
tion, and the degree of formation of soluble
pigments. The optimum temperature for the
growth of most species of Streptomyces is be-
tween 25 and 30°C. Only a few of these or-
ganisms are thermophilic. Abilities to grow
under mesophilic and thermophilic condi-
tions have been recognized as important
criteria for establishing species and even
genera of actinomycetes and other microor-
ganisms.
78 THE ACTINOMYCETES, Vol. II
The optimum reaction for the growth of
actinomycetes is pH 6.8 to 7.5. When these
organisms are grown on complex organic
media, and on many synthetic media, the
reaction usually becomes alkaline. Some
actinomycetes, however, are able to grow
at pH 4.5 to 6.5 and even at pH 3.0 to 4.5.
Such forms are not common, but the reaction
of the substrate has been recognized as a
potential diagnostic property.
On the basis of their effects on dead resi-
dues and upon living forms of life, actino-
mycetes have been grouped as saprophytes
and parasites, the latter being further
grouped into plant and animal parasites.
Thus we speak of ‘‘actinomycosis,” caused
by A. bovis and A. 7sraelzi, and “nocardio-
sis,’ caused by different species of Nocardia.
We associate S. scabies with the ‘“seab”’ of
potato tubers, and S. zpomoeae with a dis-
ease of sweet potato roots.
Genetics
Little is known about the genetic proper-
ties of actinomycetes and their possible
bearing upon problems of classification.
Certain observations have been made re-
cently, however, which offer rather promis-
ing leads in establishing species relationships.
The concept of vegetative hybridization of
Streptomyces cultures has been suggested. By
repeated growth of a culture in a sterile fil-
trate of sand-macerated mycelium of another
culture, the former undergoes morphological
and physiological changes. The significance
of this phenomenon and its potential utiliza-
tion for species characterization are still to
be elucidated. Sermonti and Spada-Ser-
monti (1956) demonstrated several types of
recombination among ‘wild’? and mutant
strains of S.
violaceoruber). It has been brought out in
Chapter 6 of Volume I that true hybrids can
be obtained by mating two different mutant
coelicolor (most probably S.
strains of an actinomycete. Welsch (1958)
suggested that mating may offer a conven-
ient criterion for the practical definition of
a species. The assumption was thereby made
that a species is distinct if it does not cross,
or gives only unfertile crosses with other
similar species.
Type Cultures
An important, and often-used technique
in species characterization of actinomycetes
is that of comparing fresh isolates with
type cultures. For the higher forms of plant
life, species characterization is facilitated by
study of preserved herbarium specimens. For
microorganisms, special collections of named
cultures are available for study. These cul-
tures allow comparisons of living material,
since dead or dried cultures are of but little
significance.
In establishing type cultures of actino-
mycetes it Is important to keep in mind the
fact that such cultures undergo considerable
variation when grown for a long time upon
artificial media. Some of the cultures may
lose their ability to produce aerial mycelium
and are thus deprived of properties of major
diagnostic value. Unknown strains of Strep-
tomyces free from aerial mycelium may even
be considered as species of Nocardia.
According to Pridham, * if reasonably fresh
isolates were maintained on the proper media
and preserved by lyophilization, the indi-
vidual laboratory would experience far fewer
difficulties than have been experienced in the
past. Pridham reported that since 1953, with
the use of these media and techniques, very
rarely have strains been found that produce
no aerial mycelium (which is generally well
sporulated) on the isolation media; a very
low incidence of strain degeneration has been
noted in active cultures as determined by
the methods of assessment, and all isolates
have been routinely lyophilized. These
lyophil tubes, opened from time to time,
have been found to give cultures that are
* Personal communication.
CHARACTERIZATION OF STREPTOMYCES SPECIES 79
Fiaure 21. Formation of straight sporophores by Streptomyces sp., X 1500 (Courtesy of Miss A.
Dietz, Dept. of Microbiology, Upjohn Co., Kalamazoo, Mich.).
equivalent to the original soil isolates, as
determined by the methods of assessment
used.
The prior growth of the organism in soil
media (sterile soil treated with a small
amount of CaCO, , if acid, and with a half
per cent of dried blood) or in carbon- or ni-
trogen-poor media, its refrigeration or its
lyophilization—each or all tend to prevent
degeneration and thus preserve the original
characteristics of the type culture. The cul-
tures that have already degenerated will
tend to regain their original properties as a
result of such treatments.
Shinobu (1958) suggested the following
method for making a soil medium: Into a
test tube of 1.5 em diameter place 7 g dried
fertile soil; add 1.5 ml of 2 per cent solution
80 THE ACTINOMYCETES, Vol. II
of glycerol; make up the water content to
about 20 to 25 per cent. Sterilize the tubes
at 20 pounds for 20 minutes. The culture
strains are inoculated on this medium and
incubated at 28-30°C. When the growth of
the organism is successful, white aerial my-
celium appears first on the surface of the
soil; when the culture matures the charac-
teristic color of the aerial mycelium is pro-
duced.
There is always the danger that an old
culture, transmitted from one laboratory to
another, may either have become modified or
have lost some of its original properties. It
may have become contaminated, and the
contaminant may eventually replace the
original culture. One must also remember
that different investigators have often based
their descriptions of a particular species not
upon the original culture but upon subse-
quent isolates, which may or may not repre-
sent the same species.
Finally, many holo-type cultures are not
available at all. Therefore, in some cases
type cultures are not reported. When re-
ported, they usually refer to the Institute of
Microbiology Collection (IMRU), the
American Type Culture Collection (ATCC),
or to the Agricultural Research Service
Culture Collection of the Northern Re-
gional Research Laboratory, U.S. Depart-
ment of Agriculture (NRRL). Other collec-
tions include Kidige Technischen Hochschule
(ETH) Zurich; Centraalbureau voor Schim-
meleultures (CBS) Baarn; and Institute of
Applied Microbiology, University of Tokyo
(IAM).
Standard Media
Some media are more favorable than oth-
ers for sporulation of Streptomyces cultures.
In view of the importance of sporulation in
characterizing a species (also in placing an
organism in the proper genus), it is essential
to select favorable media. Furthermore, since
some forms tend to lose the property of
sporulation on continued growth, special
precautions must be taken in preserving
such cultures. The loss of aerial mycelium
may be reversible or irreversible. Since non-
sporulating streptomycetes may resemble
nocardiae and since certain nocardiae have
been reported to produce aerial mycelium
and spores similar to those of typical Strep-
tomyces cultures, the element of confusion
between the two genera always exists.
Description of Streptomyces Species
It is commonly believed that to charac-
terize a species it 1s essential to describe a
large number of its morphological and phys-
iological properties. This procedure is not
always helpful, especially if based upon un-
reliable criteria. The medium may not be
readily duplicated, or conditions of growth
may be different, or the inoculum may not
be prepared in the same way. Because of
these and other variations, many cultures
recently isolated have been described as new
species. Another reason is that it is much
vasier to create a new species than to at-
tempt to correlate the characteristics of a
freshly isolated culture with those of known
species already described in the literature.
Numerous species also have been
created to facilitate the obtaining of patents.
Hesseltine ef al. (1954) suggested that the
following steps be taken in the taxonomic
hew
study of a Streptomyces species:
1. Collection of strains on
pigmentation of aerial mycelium.
2. Study of the morphology of strains
the basis of
erowlng on a number of media favorable to
sporulation.
3. Examination of the color of spores of
strains growing on optimum sporulating me-
dia. Five color groups were recognized: (a)
lavender, red, or pink; (b) blue, blue-green,
or green; (c) yellow; (d) white; (e) gray,
gray-brown, olive-gray, or dark gray.
4. Study of cultural characters of strains
on various synthetic and organic media.
CHARACTERIZATION OF
5. Analysis of certain physiological and
biochemical properties, notably action on
gelatin, starch, milk, and peptone-iron agar;
nitrate reduction; utilization of carbon and
nitrogen compounds; antibiotic action, com-
prising formation of and sensitivity to anti-
bioties.
6. Identification
known species, and preservation of cultures.
Pridham* emphasized that recently he has
been placing principal emphasis on micro-
of new strains with
morphology, secondary emphasis on chro-
mogenicity (deep brown to black diffusible
pigments), and tertiary emphasis on color of
aerial mycelium.
In an effort to determine whether freshly
isolated cultures can be identified on the
basis of published descriptions and what con-
ditions justify the creation of new species,
several obvious comparisons were made
(Waksman, 1957). Certain strains that
might be included in various important
species or group-species were critically ex-
amined. The following conclusions were
reached:
At present, various morphological, cultural,
and biochemical properties are known which make
it possible to establish definitely certain distinct
species among the actinomycetes. Some of these
characters are constant within certain conditions
of nutrition and environment, others are varia-
ble. Certain additional properties may be re-
quired in order to establish the degree of varia-
tion of a culture before it can be recognized as a
new species.
Certain categories of relationships among the
actinomycetes must be taken into consideration
in order to establish definitely the systematic
position of a given culture. These may be briefly
summarized as follows:
1. On the basis of all the accumulated evi-
dence, actinomycetes are shown to belong defi-
nitely to the bacteria.
2. The position of the true actinomycetes in
relation to related bacterial forms, notably the
* Personal communication.
STREPTOMYCES SPECIES SI
mycobacteria and corynebacteria, must be recog-
nized; this is true especially of certain nocardial
types.
3. The generic interrelationships among the
actinomycetes are highly significant. The separa-
tion of members of the genus Streptomyces from
those of Nocardia is difficult, especially when one
is dealing with nonsporulating forms of the first
and sporulating forms of the second. The recent
addition of two new genera, Actinoplanes and
Streptosporangium, and the recognition of certain
thermophilic groups as separate genera add fur-
ther problems to these generic interrelationships.
4. Within each genus, certain groups, species-
groups, or series must be recognized. A combina-
tion of morphological and cultural
permits the establishment
Some of these comprise a large number of forms
with many variable characteristics.
properties
of species-groups.
5. Differentiation of individual species within
each group is based upon a combination of cul-
tural and biochemical properties. The production
of specific antibiotics and the utilization of differ-
ent sugars are ample illustrations of this.
6. Cognizance of the strains and varieties
within each species must finally be taken. This
may be based upon certain qualitative properties,
such as sensitivity to phages, or quantitative
properties, such as production of a given anti-
biotic, vitamin, or enzyme, or sensitivity to a
given antibiotic.
The fact that a culture becomes important
for the production of a particular metabolic
product, such as an antibiotic, an enzyme,
or a vitamin, may impart to the culture
particular significance for characterization
purposes.
The existence of physiologic races or varie-
ties among species of actinomycetes, espe-
cially among those placed in the genus
Streptomyces, has been fully recognized.
Just as in improving higher forms of life
one is always faced with the selection of
varieties resistant to disease, or giving higher
yields, or having other desirable qualities,
so one must select strains of actinomycetes
on the basis of resistance to phage or of pro-
duction of higher yields of a given antibiotic
or other metabolic product.
Ghva ip tier 49
Systems of Classification and
Identification of Groups and
Species of the Genus
Streptomyces
Principles of Separation of Genera
The historical background and various
systems of classification of the order Actino-
mycetales in general and of the actinomy-
cetes in particular have been discussed in
detail in Chapter 4 of Volume I. The princi-
ples underlying the generic and specific
separation of the organisms are presented
in Chapter 4 of the present volume. The
variability and overlapping among genera
and species have been emphasized in Chap-
ter 6 of Volume I. Certain important factors
pertaining specifically to the genus Strepto-
myces must be considered before any dis-
cussion is presented of the separation of this
genus into subgenera, series (sSpecies-groups),
species, and varieties.
Among the factors that must be empha-
sized in any attempt to classify actinomy-
cetes, the following three are most impor-
tant: (a) the nature of the substrate (or
vegetative) growth and the nature of the
aerial mycelium, if any; (b) the degree of
variability of the cultures; and (c) the effect
of the composition of the medium. To facili-
tate recognition of the organisms and to
establish constant and variable differences
fol purposes, well-defined
media and standard conditions of cultiva-
tion must be used.
classification
Actinomycetes are differentiated from the
true bacteria by their filamentous growth
and by their true branching. It is often
dificult, if not impossible, to distinguish
between the profuse branching of certain
mycobacteria and the short-lived mycelium
of the nocardias, except for the fact that
the latter produce mycelium consistently
in the early stages of their development.
There is a gradual transition between the
mycobacteria and the nocardias. It also is
often difficult to differentiate between the
nocardias and the streptomyces. The latter
are characterized the constant and
marked nature of their aerial mycelium,
whereas
by
characterized
largely by the transitory and undifferenti-
ated nature of this mycelium.
the nocardias are
In establishing differences between no-
cardias and streptomyces, one must con-
sider the following factors:
1. Nocardias usually have been consid-
ered incapable of forming aerial mycelum
that could be differentiated from the sub-
strate mycelium. It also has usually been
assumed that no spirals are ever formed
the mycelium.
and Mihm
that certain nocardias are able to form aerial
from Recently, however,
(Gordon (1958) have reported
mycelium similar to that of streptomyces
GROUPS AND SPECIES OF
and that spirals also may be formed. A
streptomycete forms a characteristic aerial
mycelium. This property may be lost, how-
ever, on continued cultivation or under
special conditions of treatment. The aerial
mycelium frequently develops characteristic
spirals, tufts (Fig. 22), or verticils (whorls).
2. A streptomycete usually multiples by
the concentration and fragmentation of the
protoplasm within a filamentous cell, fol-
lowed by the dissolution of the cell mem-
brane. The fragmented portions of the my-
celium usually develop, under favorable
conditions, into fresh mycelium, either by
germ tubes or by lateral budding. Spores or
conidia are produced. The substrate my-
celium does not segment spontaneously into
bacillary or coccoid forms, but remains non-
septate and coherent even in old cultures,
thus forming the characteristic tough tex-
tured, leathery growth.
3. In nocardias, the aerial hyphae are
believed to represent an upward extension
of the substrate mycelium, and usually do
not exhibit any differentiated protoplasm.
When a streptomycete loses its capacity to
produce aerial hyphae, a form analogous to
that of a nocardia may result, except for
the structure of the mycelium that
faculty of the degenerated culture to regain
and
the lost capacity.
4. Another difference between nocardias
and streptomycetes is the acid-fastness or
partial acid-fastness of some of the forme!
when grown in certain media; the latter are
never acid-fast.
As pointed out in Chapter 1, Gordon and
Smith (1955) proposed six distinctive char-
acters for the separation of the two genera.
These criteria are: (a) colony structure; (b)
casein hydrolysis; (¢) dissolution of tyrosine
and xanthine; (d) acid production from glu-
cose and glycerol; (e) lack of acid formation
from arabinose, xylose, lactose, mannitol,
and inositol; and (f) utilization of acetate,
propionate, pyruvate, malate, and succinate.
Strains of organisms giving positive re-
GENUS STREPTOMYCES 835
Ficure 22. Tuft formation in aerial mycelium
of the Streptomyces griseus type.
sults in five or six of the physiological tests
belong to Streptomyces; strains with four
to six negative reactions, to Nocardia.
Although considered as somewhat arbi-
trary, these criteria allowed clear-cut generic
separation of 97 per cent of 251. strains
studied, regardless of their morphological
variation. Ninety-six per cent of the strains
received as Streptomyces were positive in
five or six of the following reactions: hy-
drolysis of casein, dissolution of tyrosine,
and acid production from xylose, mannose,
maltose, and lactose. Strains that no longer
formed aerial hyphae and spores, but known
ot
ones, also were positive in five or six of
the
beled Nocardia gave negative results in four
to be descendants typical sporulating
these tests. Two-thirds of strains la-
to six of the same tests. Of the remaining
third of the straims
24 had the
strains of Streptomyces and were assumed to
received Nocardia,
as
same reactions as accepted
84 THE ACTINOMYCETES, Vol. II
be mislabeled; seven were listed temporarily
as Intermediates between the two genera.
The differentiation between Streptomyces
and the other genera of actinomycetes is
not very difficult. The formation of aerial
mycelium and the manner of sporulation
are markedly distinct for Streptomyces as
compared to A/icromonospora. Species of
Thermoactinomyces also produce an aerial
mycelium, similar to that of species of
Streptomyces, but they form single spores,
similar to those of A/icromonospora. The
other thermophilic genera, as well as the
genera Waksmania (Microbispora*), Actino-
planes, and Streptosporangium also can be
differentiated from Streptomyces, as shown
in Chapters 8 to 11.
Among the numerous species belonging
to the various genera of actinomycetes,
those of the genus Streptomyces are by fai
the most important, largely because of their
wide distribution, their greater abundance,
and their ability to produce antibiotics and
vitamins and to carry out important chemi-
cal conversions. Hence a detailed considera-
tion of this genus is justified.
Description of Genus Streptomyces
Streptomyces species produce a_ well-de-
veloped mycelium. The diameter of the
hyphae seldom exceeds 1.0 u and is usually
only 0.7 to 0.8 un. The hyphae vary greatly
in length: some are long with limited branch-
ing; others are short and much branched.
The substrate mycelium does not form cross
walls; it does not break up into rod-shaped
and coccus-like bodies. Reproduction occurs
by means of spores or by bits of mycelium.
Spores or conidia are formed in_ special
spore-bearing hyphae or sporophores which
arise from the aerial mycelium either mono-
podially or in the form of tufts or verticils.
* Both designations were published, in different
journals, the same month and the same year.
Priority has not been definitely established.
The sporulating hyphae are straight or
curved. The curvatures range from mere
waviness to perfect spirals, which may be
compact, in the form of fists, or long and
open (Fig. 23).
The spores of streptomycetes comprise
four types: smooth, warty, spiny, or hairy.
About one-third of the gray- to brownish-
spored species were found (Tresner et al.,
1960) to form spiny, warty, or hairy spores;
the remainder were smooth-spored. All the
blue- to blue-green-spored forms had spiny
spores. White, vellow, cream, or buff types
had smooth-walled spores. All the pinkish-
tan-spored group had smooth spores, with
the exception of S. erythreus and S. pur-
purascens Which had spiny spores. The con-
clusion was also reached that, because of
the variation of spore size and shape, those
properties are of lmited usefulness for
taxonomic differentiation.
The growth of Streptomyces ‘‘colomies’
on artificial media is smooth or lichenoid,
hard and densely textured, raised, and ad-
hering to the medium. The colony is usually
covered completely or partially (in the form
of spots or concentric rings) by aerial my-
?
celium, which may be variously pigmented,
depending on the species and on the com-
position of medium. In liquid media, es-
pecially in shaken cultures, growth of strep-
tomyces is usually in the form of flakes,
which gradually fill the container, or in the
form of spherical growths; the former type
of growth is the more desirable from the
point of view of antibiotic production.
Many of the cultures, either in the form
of colonies on the surface of solid media or
as flaky growth in submerged culture, may
undergo rapid lysis. The production of anti-
biotics usually corresponds with the lysis
of the cultures. Frequently, the lysis is
brought about by a phage, known as actino-
phage, which exerts an injurious or destruc-
tive effect upon the mycelium.
GROUPS AND SPECIES OF GENUS STREPTOMYCES 85
FIGURE 23.
3ull. Torrey Botan. Club 82: 111, 1955).
Classification Systems of the Genus
Streptomyces
Early Systems of Classification
Although the earlier systems of classifica-
tion of actinomycetes were also supposed to
be concerned with all the forms usually in-
cluded in this group, they represented
Various strains of Streplomyces californicus (Reproduced from:
Jurkholder, P.
Re el al.
largely those forms that are now included
in the genus Streptomyces. This is true, for
example, of the first classification of San-
felice (1896), and the subsequent ones of
Krainsky (1914) Waksman
Curtis (1916). Only the more comprehensive
and of and
and more recent systems are presented
here. The earlier ones were given in Chapter
t of Volume L.
86 THE ACTINOMYCETES, Vol. II
1. WAKSMAN AND CURTIS (1916) SYSTEM
This system was based upon formation of
soluble pigments in organic media, rate of
gelatin liquefaction, and structure of aerial
mycelium.
A. Gelatin rapidly liquefied; no brown pigment.
I. Spirals formed.
1. No soluble pigment on synthetic media.
Actinomyces Rutgersensis
2. Pigment formed on synthetic media.
a. Pigment dark blue.
Actinomyces violaceus-Caesert
b. Pigment brown.
Actinomyces diastaticus
II. No spirals.
1. No soluble pigment.
a. Growth orange-red, aerial mycelium
white.
Actinomyces albosporeus
Fraure 24. Sporophores of Streptomyces sp. producing spirals, X 1500 (Courtesy of Miss A. Dietz,
Dept. of Microbiology, Upjohn Co., Kalamazoo, Mich.).
————
GROUPS AND SPECIES OF GENUS STREPTOMYCES 87
b. Growth rose-colored, aerial mycelium
rosy.
Actinomyces Fradit
ce. Growth a mixture of white and yel-
low.
al. No conidia.
Actinomyces albo-flavus
bt. Abundant conidia.
a2. Conidia rod-shaped, powdery,
gray-yellow.
Actinomyces griseus
b?. Conidia spherical and oval,
growth compact, citron-yel-
low.
Actinomyces citreus
d. Growth at first colorless, then brown
to black.
al. Aerial mycelium white.
Actinomyces alboatrus
bt. Aerial mycelium dark gray.
Actinomyces Lipmanit
. Soluble pigment produced.
a. Soluble pigment green.
bo
Actinomyces Verne
b. Soluble pigment dark blue.
Actinomyces violaceus-niger
B. Gelatin liquefied; brown pigment
formed.
I. Spirals produced.
a. Growth rose-colored; aerial mycelium
rosy.
rapidly
Actinomyces roseus
b. Growth colorless; aerial mycelium
golden brown.
Actinomyces aureus
ce. Growth slightly brown; aerial mycel-
ium white.
Actinomyces Halstedii
Il. No spirals.
1. No soluble pigment on synthetic media.
a. Growth red to red-orange; no aerial
mycelium.
Actinomyces Bobili
b. Growth white; aerial mycelium white.
al. Aerial mycelium thin, rare, net-
like.
Actinomyces reticult
b!. Aerial mycelium thick, white to
gray.
Actinomyces albus
2. Soluble brown pigment produced on
synthetic media.
a. Aerial mycelium white, abundant.
Actinomyces diastato-chromo-
genus
b. Aerial mycelium white,
late or not at all.
produced
Actinomyces
group
ce. Growth green; aerial mycelium white.
chromogenus
Actinomyces — virido-chromo-
genus
C. Gelatin slowly liquefied; no soluble pigment.
I. Spirals produced in aerial mycelium.
1. Soluble red and blue pigments.
Actinomyces violaceus-ruber
2. No soluble pigment; substrate growth
red.
Actinomyces Californicus
Il. No spirals produced in aerial mycelium.
1. Growth yellow; no soluble pigment.
Actinomyces parvus
2. Growth tends to crack; soluble brown
pigment.
Actinomyces exfoliatus
D. Gelatin slowly hquefied; brown pigment pro-
duced.
I. Spirals produced; aerial mycelium laven-
der.
Actinomyces lavendulae
II. No spirals.
1. Growth yellow; aerial mycelium gray.
Actinomyces flavus
2. Growth colorless; aerial mycelium
purplish-white.
Actinomyces purpurogenus
3. Growth black; aerial mycelium scant.
Actinomyces — erithrochromo-
genus
4. Growth purple; no aerial mycelium.
Actinomyces purpeo-chromo-
Genus
2. WAKSMAN SYSTEM (1919)
This was a modification of the previous
system and was based upon a study of 41
species. An examination was made of the
morphology of the aerial mycelium on two
media; growth, aerial mycelium, and solu-
ble pigment on 12 different media; various
biochemical properties, such as carbon and
nitrogen utilization, proteolytic activities,
diastase and invertase formation, reduction
of nitrate to nitrite, and change in reaction
of medium. A brief outline is presented here:
A. Soluble pigment produced on organic
media.
Py
ia
4
{/
(/
P qi a
- <Gi frre 7re se e
eee
ae, fey
CLIT
a jm yaya!
v. =
14
CLrechs/2°
ya gel
FIGURE 25. One of the early studies on the strueture of the aerial mycelium and the manner of sporu-
lation of Streptomyces cultures (Reproduced from: Drechsler, C. Botan. Gaz. 67: 66-83, 147-168, 1919).
SS
GROUPS AND SPECIES OF
I. Pigment deep brown.
Il. Pigment faint brown, golden yellow,
or blue.
B. No soluble pigment on organic media.
I. Strongly proteolytic.
Il. Weakly proteolytic.
The need for the recognition of species-
groups was also emphasized: ‘‘All the cul-
tures should be divided into groups, the
representatives of which have
morphological, physiological and cultural
characters. These species-groups may show
slight variations within the groups, when
common
several representatives are compared, but
all of them possess in common the main
distinguishing characters of the species, and
are distinctly different from any other
species-group.”’
3. JENSEN’S SYSTEM (1930)
Jensen modified the above system of
Waksman, in describing 90 strains of actino-
mycetes, largely streptomycetes, isolated by
him. These strains were divided into several
species-groups.
A. No pigment produced on protein media.
1. Red or blue pigments in synthetic media;
marked reduction of nitrates.
Actinomyces violaceus-ruber
to
No red or blue pigments.
a. Typical golden pigment in all synthetic
media.
Actinomyces fulvissimus
b. Pigment not typical; abundant aerial
mycelium.
al. Aerial mycelium on synthetic media
dark slate-gray; sulfur-
yellow pigments sometimes formed.
a2. Vegetative mycelium on synthetic
lemon- or
agar light colored.
Actinomyces cellulosae
b?. Vegetative mycelium on synthetic
agar turning dark.
Actinomyces olivaceus
. Aerial mycelium greenish- or yellow-
ish-gray; very rapid liquefaction of
gelatin and blood-serum.
a®. Aerial mycelium greenish.
Actinomyces griseus
GENUS STREPTOMYCES 89
b?. Aerial mycelium yellowish.
Actinomyces griseoflavus
B. Typical brown pigment in protein media
(“chromogenus’’ species).
1. Deep brown growth and pigment in all syn-
thetic media.
Actinomyces phaeochromo-
genus
2. Pigment in synthetic media of other color or
absent.
a. Aerial mycelium absent or in
typical red vegetative mycelium.
traces;
Actinomyces bobili
b. Aerial mycelium more or less abundant.
al. Typical
agar.
red pigment in synthetic
Actinomyces — erythrochromo-
genus
. Pigment not red.
a®, Aerial mycelium rose to cinna-
mon-brown.
Actinomyces roseus
b?. Aerial mycelium abundant, char-
acteristic lead-gray; hght brown
pigment in synthetic media.
Actinomyces diastatochromo-
genus
Jensen
term
contribution,
the
actinomycetes
subsequent
(1931) emphasized again that
“species” applied — to
should be used in the sense of Waksman’s
“species-groups,” ora “broad group of strains
in
In a
as
agreeing celtain outstanding morpho-
logical and biological features’; otherwise,
ce . . . on .
every strain of actinomycetes isolated from
a plating from an ordinary soil could then
be raised to the rank of species.”
4. KRASSILNIKOV SYSTEM (1941)
This system was based primarily upon
the morphology of sporophores of the cul-
tures and the shape of their spores, and
secondarily upon the pigmentation of the
cultures.
A. Sporophores branching monopodially.
I. Spiral-shaped sporophores, produced on
hyphae of aerial mycelium.
1. Spores spherical or oval.
a. Cultures colorless, not producing any
pigmentation.
90
b.
Cc.
d.
e.
THE ACTINOMYCETES,
al. Aerial mycelium white.
a®, Saprophytes, hving on dead
material.
Actinomyces albus
b2. Parasites, living on plants.
Actinomyces totschidlowskii
bt. Aerial mycelium dark gray.
Actinomyces griseus
cl, Aerial mycelium green.
Actinomyces glaucus
Cultures pigmented blue.
a'. Pigment of the anthocyanin type,
similar to litmus.
Actinomyces coelicolor
bt. Blue pigment not changing with
acidity of medium.
Actinomyces cyaneus
Cultures violet, forming two basic
pigments (red and blue), both dis-
solved into the substrate.
al. Cultures not forming any fluores-
cent substance in liquid media.
Actinomyces violaceus
b!. Cultures producing in synthetic
media a fluorescent substance of
blue-green color similar to pyo-
eyanin.
Actinomyces pluricolor
Cultures black-violet, forming red
and blue pigments, as well as a brown
pigment of the type of melanin, which
changes the violet color of the culture
to violet-black.
Actinomyces violaceus-niger
Cultures red-colored, producing pig-
ments insoluble in water, of the hpo-
actinochrome type; color of medium
not changing with acidity.
al. Cultures not forming any brown
or black pigments; they are al-
sometimes with a
brownish tinge, but not black.
ways red,
Actinomyces ruber
bt. Cultures producing on synthetic
media, in addition to pigments, a
black or dark brown substance
which gives the culture a red-
brown to black color.
Actinomyces melanocyclus
Cultures yellow,
brownish-yellow.
citron-yellow, or
al. Saprophytes.
Actinomyces flavus
b!. Living on plants.
Actinomyces setonii
i
lave
Vol. II
Cultures orange.
al, Saprophytes.
Actinomyces aurantiacus
b!. Parasites.
Actinomyces phenotolerans
Cultures green or brownish-green.
Actinomyces viridochromo-
genes
Cultures black, producing a pigment
of the melanin type.
Actinomyces niger
Cultures pigmented dark brown, but
not black.
al, Saprophytes.
Actinomyces chromogenes
b!. Plant parasites.
Actinomyces gracilis
2. Spores cylindrical or elongated.
a.
loys
—
al
Cultures colorless.
Actinomyces longisporus
Cultures red, mostly
straight.
al. Saprophytes.
Actinomyces
sporophores
longis porus -
ruber
b!. Parasites.
a2. Living in bodies of men and
animals.
Actinomyces spumalis
2. Living on plants.
Actinomyces salmonicolor
Y
Ss
Cultures orange.
Actinomyces fradiae
. Cultures yellow.
al. Saprophytes.
Actinomyces
flavus
b!. Parasites living on plants.
longisporus-
Actinomyces scabies
Cultures citron-yellow.
Actinomyces virgatus
Cultures green.
Actinomyces viridans
Cultures brown or chocolate-colored.
Actinomyces halstedit
Cultures black.
Actinomyces nigrificans
Sporophores straight or wavy, but not
spiral.
1. Spores produced by means of fragmenta-
tion of plasma within cells.
a.
Spores spherical or oval.
al. Cultures colorless.
Actinomyces globisporus
GROUPS AND SPECIES OF GENUS STREPTOMYCES 91
b!. Cultures green.
a®. Saprophytes.
Actinomyces viridis
b2. Plant parasites.
Actinomyces cretaceus
e!. Cultures brown.
Actinomyces globosus
b. Spores cylindrical or elongated.
alt. Cultures colorless.
Actinomyces candidus
b'. Cultures pigmented.
Actinomyces cylindrosporus
2. Spores produced by means of segmenta-
tion of aerial hyphae.
a. Cultures colorless.
Actinomyces farinosus
b. Cultures pigmented red.
Actinomyces oidiosporus
Actinomyces rectus
~
. Cultures vellow-orange.
Actinomyces longissimus
d. Cultures blue.
Actinomyces caeruleus
. Cultures brown.
Actinomyces fumosus
a)
B. Sporophores produced in verticils.
I. Sporophores straight.
Actinomyces verticillatus
Il. Sporophores spiral-shaped.
L. Spores spherical, oval.
Actinomyces reticuli
Actinomyces reticulus-ruber
2. Spores cylindrical, elongated.
Actinomyces circulatus
Detailed consideration was given to some
of the larger groups of the genus, notably
to the Albus and Flavus groups, as will be
shown in Chapter 6.
5. WAKSMAN AND HENRICI SYSTEM (1943)
This system was used in the last two edi-
tions of Bergey’s Manual of Determinative
Bacteriology (1948, 1957). It may be listed
here among the earlier was
based primarily upon the ecology of the
systems. It
organisms, production of soluble pigments
in organic and synthetic media, and proteo-
lytic properties.
A. Saprophytes; psychophilic to mesophilic.
1. Soluble pigment on organic media
other than brown, or faint brown.
a. Pigment absent or faint brown
only.
b. Pigment blue.
c. Pigment at first
brown, ete.
green, becoming
2. Soluble pigment on organic media
brown.
3. No soluble pigment produced in or-
ganic media.
a. Proteolytic action strong.
b. Proteolytic action limited.
c. Proteolytic action very weak.
B. Saprophytes; thermophilic.
1. Yellowish
static.
2. Dark-colored
potato; nondiastatic.
growth on potato; dia-
abundant growth = on
5}. Thermotolerant cultures.
C. Plant parasites or cultures isolated from
diseased plants or from soil in which
diseased plants were grown.
1. Isolated from potato scab or from
soil in which scabby potatoes were
erown.
bo
Grown on or isolated from sweet
potatoes.
3. Isolated from scab on mangels and
sugar beets.
D. Isolates the
animal body, hyphae often show clavate
from animal tissues; in
enlargements at the ends.
1. Limited proteolytic action.
2. Strong proteolytic action.
—
[ter
Produce only substrate growth and no
aerial mycelium.
The last system, like the earlier ones, may
now be considered as of purely historic in-
terest. For the purpose of this treatise, it
has been considerably modified. It has been
greatly enlarged to include all newly de-
scribed organisms belonging to the genus
Streptomyces. Some of the features of the
older system, notably those pertaining to
ecology, have been left out altogether.
92 THE ACTINOMYCETES, Vol. II
Recent Systems of Classification
Several systems for classifying species
belonging to the genus Streptomyces have
recently been proposed. Some of these have
been selected for detailed examination. They
are based largely upon morphology, cultural
and biochemical properties, or combina-
tions of these.
1. HESSELTINE, AND PRIDHAM
SYSTEM (1954)
BENEDICT,
In this system, emphasis was laid upon
morphology as the basis for separation of
the genus into five basic groups. After a
study of hundreds of cultures on a variety
of media, the conclusion was reached that
the morphology of any one strain of Strepto-
myces is essentially the same on any medium
where sporulation occurs. The major groups
were subdivided into a number of subgroups
on the basis of cultural properties, pigmen-
tation of spores, and other criteria.
I. Sporophores not restricted in length,
bearing fertile branches in verticils,
with spores more or less strongly at-
tached.
1. Fertile branches in simple verticils,
branches not ending in spirals.
Fertile branches in simple verticils,
branches ending in spirals.
3. Fertile branches with compound
verticils, branches not ending in
bo
spirals.
II. Sporophores with branches all straight,
never ending in spirals; verticils ab-
sent.
III. Sporophores predominantly in tufts,
never verticillate; outline of branches
flexuous and irregular.
IV. Sporophores with branches ending in
absent; sporo-
stalks bearing
as short stalks
spirals, verticils being
phores either as long
very short branches, or
bearing branches irregularly.
1. Branches ending in open spirals with
many turns.
2. Branches ending in closed spirals
with few turns, thus appearing as
tight knots.
V. Sporophores with long and_ straight
branches with spirals of large diameter
at their ends; spirals usually with only
a few turns, never verticillate.
No strains were observed in which the
sporophores were unbranched, except when
they were growing under unfavorable con-
ditions or where degenerated type cultures
were studied.
Seven major Streptomyces groups were
thus created as indicated by the following
key:
I. Sporophores produce verticils; spores not
readily separating; aerial mycelium white,
pink, lavender, or tan.
Group |. Streptomyces reticuli
1. Verticil branches simple.
a. Sporophores straight.
b. Sporophores spiral-shaped.
2. Verticil branches compound.
a. Ultimate branches straight.
b. Ultimate branches spiraled.
II. Sporophores not producing any verticils;
spores readily separate; color of aerial myce-
lium often not pink, white, lavender, or tan.
1. Spirals always formed; color of aerial my-
celium blue, blue-green, or green.
Group II. Streptomyces viridochro-
mogenes
2. Spirals may or may not be formed; color of
aerial mycelium different.
a. Spirals never formed, tufts often present ;
color of aerial mycelium greenish-tan or
tan, never white.
Group III. Streptomyces griseus
b. Spirals produced; color of aerial myce-
lium lavender, red, pink, or nearly tan.
Group IV. Streptomyces lavendulae
Sporophores straight.
a. Spores white or nearly so.
vw
Group V. Streptomyces albus
b. Aerial mycelium never white.
al. Aerial mycelium yellow.
Group VI. Streptomyces parvus
bt. Aerial mycelium gray, gray-brown,
olive-gray, blackish-gray.
Group VII. Gray-spored group.
It was suggested that the last group could
GROUPS AND SPECIES OF GENUS STREPTOMYCES 93
be subdivided on the basis of spiral forma-
tion.
2. FLAIG AND KUTZNER SYSTEM (1954, 1960)
Flaig and Kutzner (1954) and Kutzner
(1956) studied about 2000 Streptomyces cul-
tures, 63 of which were authentic species
and the rest fresh soil isolates. They charac-
terized their cultures by several criteria in-
cluding cultural characteristics on complex
and synthetic media, morphology of the
aerial mycelium, physiological properties,
and antibiotic activity against five test or-
ganisms. On the basis of these studies, a
key was prepared. At first the material was
divided into ‘“‘groups’? on the basis of the
color of the aerial mycelium on oatmeal
agar, six spore colors being recognized. The
gray color group was further divided, on the
basis of the color of the substrate mycelium
and soluble pigment on this medium, thus
resulting in the 10 groups shown in Table 9.
Later, however, Flaig and Kutzner (1960)
reached the conclusion that the subdivision
of the gray-spored color group may lead to
difficulties when new isolates have to be
placed in one of these groups, although this
system proved to be quite useful in studying
many strains at the same time.
Each group was further divided into sub-
groups (altogether 382) on the basis of pig-
ment formation on glucose-peptone agar,
morphology of the aerial mycelium, anti-
biotic activity against five test organisms,
and cultural several
media. The shape of the spores (observed
with electron microscope) was given for 175
of the 382 subgroups. Various authentic
characteristics on
species were included in these subgroups
according to their characteristic properties.
The following species were placed in the
various groups.
Group I: S. albus, S. griseus, S. chryso-
mallus, S. coelicolor, S. cali-
fornicus
Group II: — S. longispororuber, S. bobiliae,
S. roseochromogenes, S. vene-
zuelae, S. phaeochromogenes
Group III: 8S. lavendulae, S. xanthophaeus
Group IV: S. flavogriseus, S. globisporus,
S. flaveolus
Group V: S. diastaticus, S. globosus
Group VI: S. flavus
Group VII: S. craterifer, S. griseolus, S.
halstedii, S. hygroscopicus, S.
aureofaciens, S. violaceoruber
Group VIII: S. purpurascens
Group IX: S.
genes, S. chartreusis
Group X:_ iS.
prasinopilosus
cyaneus, S. viridochromo-
hirsutus, S. prasinus, S.
TABLE 9
Grouping of the genus Streptomyces according to characteristic coloration (Flaig and Kutzner, 1960)
Group Aerial mycelium Substrate mycelium and soluble pigment
I Yellowish to yellow-gray Colorless, brownish, reddish, greenish
II Light rose to reddish Colorless, orange, greenish, brownish to
g
dark brown, pink to dark red
IIL Gray-rose (lavender) Colorless, orange, brownish to dark brown
i io
IV Light gray to gray Yellowish-green to green
. aS . S S
V Gray Brown
VI White to gray (cottony) Colorless, brownish to brown
VII Gray (dusty) Colorless, greenish-gray, brownish-gray,
2 : g era) Bra)
orange-brownish, red or blue violet
VIII Light gray or pink Violet
IDS Blue Blue-purple, bluish-green, brownish
pur}
x Green Colorless, greenish, brownish to rose-red
94 THE ACTINOMYCETES, Vol. II
3. SYSTEM OF YAMAGUCHI AND SABURI (1955)
Yamaguchi and Saburi also used morpho-
logical features as the primary basis for the
separation of the genus Streptomyces into
groups, and physiological characteristics
for further separation. Although they were
concerned primarily with species of Strepto-
myces possessing antitrichomonal properties,
their system may apply to the genus as a
whole.
I. Sporophores straight, tuft-forming tendency
in the margin; no verticils or spirals.
1. Aerial mycelium gray.
a. Soluble pigment on protein media.
Light purple, reddish-purple, purplish-
brown, sometimes yellowish-brown.
Streptomyces purpeofuscus
b. No soluble brown pigment.
Streptomyces fasciculus
Aerial mycelium pale yellowish-green.
a. Soluble pigment on protein media
brown—ATCC Culture No. 3309.
b. Soluble pigment brown.
Streptomyces griseus
II. Sporophores straight, verticils produced.
1. Cottony aerial mycelium white, light tan,
or pale pink.
a. Soluble pigment on
brown.
bo
protein media
Streptomyces reticult
b. No soluble brown pigment on protein
media.
Streptomyces hachijoensis
III. Sporophores spiral-shaped.
1. Predominantly closed spirals produced.
a. Aerial mycelium gray.
al. Soluble brown pigment on protein
media.
a2. Growth yellowish-
brown, brown, or deep brown.
colorless,
Streptomyces olivochromogenes
b?. Growth light purple to purplish-
black.
Streptomyces — purpureochro-
mogenes
b!. No soluble brown pigment.
Streptomyces aureofaciens
b. Aerial mycelium pale pink.
al. Soluble pigment on protein media
brown.
Streptomyces lavendulae
b!. No soluble brown pigment.
Streptomyces fradiae
i)
. Predominantly open spirals or compact
spirals produced.
a. Aerial mycelium white.
al. Soluble pigment on protein media
brown.
a®. Growth reddish.
Streptomyces ruber
bt. No soluble brown pigment.
Streptomyces farinosus
b. Aerial mycelium gray.
al. Soluble pigment on protein media
brown.
a®, Abundant compact spirals pro-
duced on aerial hyphae.
Streptomyces naganishir
b?. Growth colorless, white, light
yellow, or yellowish-brown.
Streptomyces diastatochromo-
genes
c?. Growth colorless, reddish-or-
ange, or reddish-purple.
Streptomyces griseoruber
bt. No soluble brown pigment.
a®. Growth colorless to creamy.
Streptomyces albus
b?. Growth colorless, white, light
yellow, or yvellowish-brown.
G 167
myces Cacaor)
ce. Growth colorless, light yellow,
or ight pinkish-brown.
(resembling Streplo-
Streptomyces albogriseolus
d?. Growth purple to pink to red.
Streptomyces californicus
ce. Aerial mycelium gray, but on certain
media with dark, glistening
patches.
moist
Streptomyces hygroscopicus
IV. No characteristic features of aerial hyphae.
1. Very limited aerial mycelium production
on various media.
a. Soluble pigment on protein media deep
brown.
al. Growth yellow to yellowish-brown.
Streptomyces flavochromogenes
b. Soluble pigment faint yellowish-brown.
Streptomyces thioluteus
2. Aerial mycelium white.
a. Growth colorless, light yellow, or red-
dish-orange.
Streptomyces ruber
4, BALDACCI SYSTEM (1956, 1958, 1959)
Following the example of Sanfelice and
Waksman, Baldacci divided the genus
GROUPS AND SPECIES OF GENUS STREPTOMYCES
Streptomyces into sections, based upon the
color of the substrate mycelium. Kach sec-
tion was divided into series, on the basis of
the color of the aerial mycelium. Each series
was divided into species.
The genus was characterized by the pres-
ence or absence of spores, the arrangement
of spores, and the ramification and breaking
up of the substrate mycelium. The species
were characterized by enzymatic reactions,
antibiotic activity, and soluble pigments
spreading through the substratum, depend-
ing on nutrition and pH.
Although Baldacci discussed the genus
under the name ‘‘Actienomyces,” he actually
meant Streptomyces, without recognizing it
as such, since he made no mention of other
species and other genera.
A. Actinomycetes cum sporophora_ solitaria
vel congregata.
Section I. Substrate mycelium
scant development on agar, showing a
cobweb-like appearance; limited
colorless;
veiled
sporulation.
1. Aerial mycelium white.
Series Albus
2. Aerial mycelium sea-green.
Series Griseus
3. Aerial mycelium green-azure.
Series Viridis
4. Aerial mycelium azure.
Series Caeruleus
5. Aerial mycelium white-wine-lavender.
Series Lavendulae
6. Aerial mycelium light pink.
Series Ioseus
~J
Aerial mycelium gray.
Series Diastaticus
II. Substrate
development generally abundant on agar
Section mycelium colored;
(creamy, lichenoid, ete.); delayed or par-
tial sporulation.
(a) Substrate mycelium yellow to yellow-
brown.
9. Aerial mycelium white.
Series Albidoflavus
(b)
(d)
(e)
95
10. Aerial
pink).
mycelium pink (white to
Series Roseoflavus
11. Aerial mycelium yellow
spots.
with gray
Series Flavus
12. Aerial mycelium grayish.
Series Aureus
Substrate mycelium yellow to
vellow.
ereen-
13. Aerial mycelium gray-white.
Series Flavoviridis
14. Aerial mycelium white to lemon-
vellow.
Series Virgatus
Substrate mycelium yellow with green
and pinkish spots.
15. Aerial mycelium white to pink.
Series WJadurae
Substrate mycelium brown to black.
16. Aerial mycelium white to gray.
Series Scabies
17. Aerial mycelium red.
Series Roseochromogenes
18. Aerial mycelium yellow.
Series Sulphureus
19. Aerial mycelium gray.
Series A ntibioticus
Aerial mycelium grayish-flesh-col-
ored.
Series Griseoincarnatus
Vegetative mycelium brown to green-
brown.
21. Aerial mycelium gray.
Series [ntermedius
Vegetative mycelium brown.
22. Aerial mycelium white to leather-
brown.
Series Rimosus
Vegetative mycelium orange.
23. Aerial mycelium seashell pink.
Series Fradiac
Vegetative mycelium flesh-rose.
24, Aerial mycelium white.
Series Bostroemi
Vegetative mycelium red.
96 THE ACTINOMYCETES, Vol. II
25. Aerial mycelium white to pink.
Series Albosporeus
26. Aerial mycelium ash-gray.
Series Cinereo-ruber
(j) Vegetative mycelium violet-blue-red.
. White to gray aerial mycelium with
different shades.
_
27
Series Violaceus
B. Actinomycetes cum sporophora opposita
et verticillata*
Section I. Substrate mycelium colorless.
1. Aerial mycelium white or whitish.
Series Circulatus
2. Aerial mycelium gray and pinkish.
Series Griseocarneus
Section IT. Vegetative mycelium colored.
(a) Substrate mycelum lemon-yellow-
creamy-colored.
3. Aerial mycelium cinnamon-colored.
Series Cinnamoneus
(b) Substrate mycelium brownish-yellow.
4. Aerial mycelium gray.
Series Reticulr
(c) Substrate mycelium brown.
5. Aerial mycelium greenish-gray.
Series Verticillatus
(d) Substrate mycelium red.
6. Aerial mycelium pinkish-red.
Series Rubrireticuli
5. GAUSE ET AL. SYSTEM (1957)
Gause et al. modified Baldacci’s system,
leaving out the sections and combining the
pigmentation of the aerial mycelium with
that of the substrate growth for series char-
acterization. Each series was subdivided, on
the basis of formation of a soluble pigment
in a complex organic medium, or of the
structure of the sporophores, or of the pig-
mentation of a synthetic medium. These
investigators, like Baldacci, adhered to the
genus designation Actinomyces, without,
however, considering the accumulated in-
concerning all other
formation genera.
* These sections were placed by Baldacci in a
separate genus ‘‘Streptoverticillium.”
Little consideration was given to previously
hamed species.
I. Aerial mycelium rose-purple,
strate mycelium colorless.
Series Lavendulae-roseus
II. Aerial mycelium rose-colored; sub-
strate mycelium yellow.
Series Fradiae
sub-
III. Aerial mycelium rose-colored; sub-
strate mycelium brown.
Series Fuscus
IV. Aerial mycelium light rose; substrate
mycelium violet:
Series Roseoviolaceus
V. Aerial mycelium rose-colored; sub-
strate mycelium red.
Series Ruber
VI. Aerial mycelium yellowish-green or
cream-colored; substrate mycelium
colored or colorless.
Series Helvolus
VII. Aerial mycelium white; substrate
mycelium colorless.
Series Albus
VIII. Aerial mycelium white; substrate
mycelium red or brown.
Series Albosporeus
IX. Aerial mycelium blue or greenish-
blue; substrate mycelium colorless or
blue-colored.
Series Coerulescens
X. Aerial myceluum = gray;
mycelium colorless.
Series Griseus
XI. Aerial mycelium gray, then black
(result of autolysis); substrate my-
substrate
celium colorless.
Series Nigrescens
XI. Aerial mycelium gray; substrate my-
celium yellow or orange.
Series Aureus
Aerial mycelium gray; substrate my-
celium yellow-brown.
Series Chrysomallus
Aerial mycelium = gray;
mycelium brown-black.
XIII.
substrate
EV
Series Chromogenes
GROUPS AND SPECIES OF GENUS STREPTOMYCES 97
Aerial mycelium gray; substrate my-
celium blue-violet or red-brown.
Series Violaceus
6. PRIDHAM, HESSELTINE, AND
SYSTEM (1957, 1958)
In their earlier system, these investigators
divided the genus Streptomyces into seven
groups. Each group was characterized by a
distinct morphology of the sporophores in
mature cultures, and by a distinct color of
the aerial mycelium. This system subse-
quently was revised. Morphological sections
and color series were established and, on the
basis of literature study and laboratory in-
vestigations, many species and known anti-
biotic-producing strains were cataloged. It
was suggested that evaluation of the compo-
nent strains in the sections and series, by
physiological tests, would allow the deter-
mination of ranges of variation and a more
logical approach to speciation in the genus.
The placement by these investigators of
strains in morphological sections, regardless
of species designation, has suggested synon-
ymy, as well as misidentification of many
strains.
The following bases were considered in
justifying these subdivisions:
1. The morphology of the sporophores of
a particular strain does not appreciably
change on substrata that support optimal
formation of aerial mycelium, sporophores,
and spores. Morphological patterns exhibited
by streptomycetes are not subject to con-
siderable variation, unless degeneration of a
particular strain has occurred through im-
proper maintenance. Morphological exami-
nations should be made after two weeks’
incubation at 28-30°C on several appropriate
media.
2. The color of the sporulating aerial my-
celium of a given strain at maturity was said
not to differ appreciably from medium to
medium. Each morphological section of the
genus can be further subdivided into color
“series.’? Each color series can be subdivided,
BENEDICT
on the basis of physiological criteria, into
‘species.”’ Additional delineation can then
be used to create ‘“‘varieties”’ or “physiolog-
if need be.
present
ical forms,”
3. The the
Streptomyces is interpreted rather broadly.
Some of the strains identified as members of
concept of
genus
the genus may in reality belong to other
genera.
The proposed sections were designated as
follows:
I. Rectus-flexibilis
straight,
spirals. Type species S.
Il. Retinaculum-apertum
(RF).
flexuous, or
Sporophores
fascicled; no
Griseus.
(RA).
phores in the form of hooks, open loops,
or greatly extended — spirals.
species S. fradiae.
Sporo-
Type
III. Spzra (S). Sporophores either short and
gnarled, or in the form of compact
spirals or of extended long and open
spirals. Type species SS. viridochro-
mogenes (lig. 26).
IV. Monoverticillus (MV). Sporophores in
the form of primary verticils attached
to long, straight branches; no spirals.
V. Monoverticillus-spira (MYV-S).
phores as primary verticils attached to
Sporo-
long, straight branches; elements of
verticils spiraled.
VI. Biverticillus (BV). Sporophores as com-
pound verticils attached to long,
straight branches; no spirals. Type
species S. cinnamomeus ft. cinnamomeus.
Biverticillus-spira — (BIV-S).
phores as compound verticils attached
to long, straight branches; elements of
secondary verticils spiraled.
In addition to the above sections, another
section was set up to include strains for
Vat:
Sporo-
which no micromorphological data were
available.
Each ‘section’? was subdivided into
“series”? based on the color of sporulating
aerial mycelium at maturity. The proposed
series were designated as follows:
1. White.
98 THE ACTINOMYCETES,
Wolk ant
Fraure 26. Spiral formation by Streptomyces 240 (Reproduced from: Naganishi, H. and Nomi, R.
J. Fermentation Technol. 32: 492, 1954).
2. Olive-buff (buff to tan to olive-buff).
3. Yellow.
. Blue (blue to blue-green to green).
5. Red (pink to red to lavender to laven-
der-gray).
6. Gray (light gray to mouse-gray to
brown-gray to gray-brown).
An additional ‘unknown’ series was set
up to include strains for which no color data
i
were available.
7. ETTLINGER, CORBAZ, AND HUTTER SYSTEM
(1958)
Ettlinger ef al. considered four major
characters of Streptomyces that were stable
and reliable enough to justify their svstem of
classification. These characters were: (a)
morphology of the spores, (b) color of aerial
mycelium, (¢) morphology of aerial myce-
lium, and (d) formation of melanoid pig-
ment.
These investigators suggested combination
of sections 4 and 6, and sections 5 and 7 of
the Pridham et al. (1958) system, since they
had never observed nonbranching verticils.
They recognized a total of 15 morphological
types distributed among the five sections.
They also recognized the following color
groups for the aerial mycelium: (1) niveus
(snow-white), (2) (yellowish- to
greenish-gray), (3) azureus (sky-blue), (4)
cinnamoneus (light carmine to brownish),
(5) cinereus (ash-gray), (6) prasinus (leek-
green).
They observed certain constant relation-
ships among some of the four basic prop-
erties. The griseus and cinnamoneus color
Griseus
groups were found to occur only in strains
with smooth spores. The azureus and pra-
sinus color groups occurred only in strains
with spiny or hairy spores. The latter always
were found associated with the occurrence
of spirals.
Other properties, such as soluble pigment
on synthetic media, antibiotie activity, and
pigmentation of substrate mycelium, were
found to be variable. Gelatin liquefaction,
milk coagulation, starch hydrolysis, and
other physiological properties were not con-
sidered of great value from a systematic
point of view, since no true negative gelatin
liquefaction or negative starch hydrolysis
was ever detected.
On the basis of the above properties, the
GROUPS AND SPECIES OF GENUS STREPTOMYCES 99
following system of classification Was pro-
posed for the genus Streptomyces:
A. Spores spiny or hairy.
I. Aerial mycelium blue.
1. Streptomyces virido-
chromogenes
II. Aerial mycelium not blue.
1. Aerial mycelium white.
2. Streptomyces pur-
purascens
2. Aerial mycelium not white.
a. Aerial mycelium green.
al. Spores with short spines.
3. Streptomyces pra-
sinus
b!. Spores with longer spines or with
hair.
a®. Spores with stiff spines.
4. Streptomyces hirsu-
tus
b?. Spores with flexible hair.
5. Streptomyces
sinoptilosus
b. Aerial mycelium gray.
al. Sporophores in verticils.
a®. Sporophores in open spirals;
no melanin formation.
6. Streptomyces nour-
sev*
pra-
b?. Sporophores in closed spirals;
melanin produced.
7. Streptomyces echi-
natus*
b!. Sporophores not in verticils.
a®. Sporophores in closed spirals.
8. Streptomyces albo-
griseolus
b?. Sporophores in open spirals.
a®. Spirals irregular.
9. Streptomyces mac-
ros poreus
bs. Spirals regular.
a', Spores spiny.
10. Streptomyces grise-
oflavus
b*. Spores hairy.
a°. Spirals with >5 turns;
melanin-positive.
11. Streptomyces pilo-
SUS
* The verticillate nature of these organisms is
open to question.
FIGuRE 27. Sporogenous coiled hyphae of Strep-
tomyces T 3110; taken from a gray area of a colony
(Reproduced from: Duggar, B. M. ef al. Ann.
N. Y. Acad. Sci. 60: 71-85, 1954).
b®. Spirals with <5 turns,
melanin-negative.
12. Streptomyces flaveo-
lus
B. Spores smooth.
I. Aerial mycelium yellowish- to greenish-
gray.
1. Melanin-negative.
13. Streptomyces — gri-
seus
2. Melanin-positive.
14. Streptomyces michi-
ganensis
Il. Aerial mycelium white.
1. Sporophores in verticils.
15. Streptomyces rubri-
reticult
2. No verticils produced.
a. Sporophores form spirals.
16. Streptomyces nive-
oruber
b. Sporophores straight or wavy.
al. Melanin-negative.
17. Streptomyces fulvis-
STMUS
b!. Melanin-positive.
18. Streptomyces
phaeochromogenes
LOO THE ACTINOMYCETES, Vol. II
IIT. Aerial mycelium hght carmine to brown-
ish.
1. Sporophores in verticils.
a. Sporophores straight or wavy.
19. Streptomyces ne-
tropsis
b. Sporophores in spirals.
20. Streptomyces tendae
2. Sporophores not in verticils.
a. Sporophores straight.
21. Streptomyces vene-
zuelae
b. Sporophores in spirals.
al, Spirals at end of long, straight
sporophores; melanin-positive.
22. Streptomyces laven-
dulae
b!. Spirals different; melanin-nega-
tive.
a®. Spirals closed.
23. Streptomyces — vio-
laceoniger
b?. Spirals open.
a’. Spirals regular, usually
>5 turns.
24. Streptomyces — fra-
diae
b*. Spirals irregular, usually
<5 turns.
25. Streptomyces ery-
thraeus
IV. Aerial mycelium ash-gray.
1. Sporophores in verticils.
26. Streptomyces — reti-
cult
2. Sporophores not in verticils.
a. Sporophores straight or wavy.
&
FIGURE 28. Sporogenous coiled hyphae of Streptomyces T 3110; taken from a blue sector of a colony
(Reproduced from: Duggar, B. M. et al. Ann. N. Y. Acad. Sci. 60: 71-85, 1954).
GROUPS AND SPECIES OF
al, Sporophores sympodially
branched.
27. Streptomyces — viri-
dogenes
. Sporophores monopodially
branched.
2. Sporophores as side branches
of sterile hyphae.
28. Streptomyces ramu-
es)
«
c
losus
2. Sporophores different.
a’, Melanin-negative.
29. Streptomyces oliva-
ceus
b’. Melanin-positive.
30. Streptomyces
bioticus
b. Sporophores in spirals.
al. Spirals closed.
31. Streptomyces hygro-
SCOptcus
antt-
b!. Spirals open.
a®. Spirals irregular, usually >5
turns.
32. Streptomyces aureo-
faciens
b?2. Spirals regular, <5 turns.
a’. Melanin-negative.
33. Streptomyces — par-
vullus
bs. Melanin-positive.
34. Streptomyces gali-
laeus
8. SHINOBU SYSTEM (1958b)
Shinobu proposed the following system
pro} £ Sy}
for grouping of the species of the genus
Streptomyces:
Group I. Monopodial branching, straight or wavy
aerial mycelium; never producing spirals.
Subgroup 1. Tyrosinase reaction: positive; ni-
trite production: positive.
Streptomyces olivaceus
~
positive ;
nitrite production: negative.
Streptomyces phaeopurpureus
Subgroup 2. Tyrosinase reaction:
Ow
reaction: negative;
nitrite production: positive.
Streptomyces sp. No. 2
Tyrosinase reaction: negative;
nitrite production: negative.
Streptomyces sp. No. 232
Group IL. Spiral formation; long or short, loose or
compact, and open or closed spirals.
Subgroup 3. Tyrosinase
Subgroup 4.
‘ GENUS STREPTOMYCES 10]
Subgroup 5. Tyrosinase reaction: positive;
nitrite production: positive.
Streptomyces viridochromo-
genes
Subgroup 6. Tyrosinase’ reaction: positive;
nitrite production: negative.
Streptomyces sp. No. 2076.
reaction:
. Tyrosinase negative;
“I
Subgroup
nitrite production: positive.
Streptomyces sp. No. 236
negative;
nitrite production: negative.
Streptomyces scabies
Subgroup 8. Tyrosinase reaction:
Verticil formation; primary and
verticils; rarely one tertiary verticil
Group III.
secondary
(Fig. 29).
Subgroup 9. Tyrosinase reaction: positive;
nitrite production: positive.
Streptomyces hiroshimensis
Tyrosinase reaction: positive;
nitrite production: negative.
Streptomyces luteoverticillatus
Tyrosinase reaction: negative;
nitrite production: positive.
Subgroup 10.
Subgroup 11.
Streptomyces roseoverticilla-
tus
reaction: negative;
nitrite production: negative.
Streptomyces olivoverticillatus
Group IV. Intermediate group of Nitella-type and
Anitella-type verticil.
Subgroup 13.
Subgroup 12. Tyrosinase
Streptomyces spiroverticillatus
Shinobu further emphasized that in the
identification of species of Streptomyces other
characteristics should be considered. These
are the following:
1. Morphological properties: type of colony,
shape of spiral, etc.
2. Physiological
nase, and amylase reactions; utilization of carbon
properties: cellulase, man-
and nitrogen sources, etc.
3. Cultural properties: growth of colony, pro-
duction of pigment, ete.
9. FROMMER SYSTEM (1959)
This system does not apply to the genus
Streptomyces as a whole but only to the
actinomycin-producing species.
A. Chromogenic group.
I. Spirals not formed on aerial mycelium.
Occasionally a few spirals are found.
102
THE ACTINOMYCETES, Vol. II
Ficure 29. Verticil formation (Nitella type) including both primary and secondary verticils (Re
produced from: Shinobu, R. Mem. Osaka Univ. Lib. Arts and Ed. B. Nat. Sei. 7, 1958).
1. Aerial mycelium on_ synthetic
media gray. Tyrosinase-negative.
Streptomyces antibioticus
mycelium on
agar
bo
Aerial synthetic
media yellow. Tvrosinase-positive.
agar
Streptomyces michiganensis
Il. Numerous spirals produced on aerial my-
celium.
1. Yellow or yellow-green pigment
duced on synthetic agar.
pro-
Streptomyces galbus
2. Soluble pigment on synthetic agar, dark
brown.
Streptomyces lanatus
B. Nonchromogenic group.
I. No spirals on aerial mycelium.
1. Strongly proteolytic. Aerial mycelium
on synthetic agar white, yellow, or
greenish.
Streptomyces chrysomallus
2. Weakly proteolytic. Aerial mycelium on
synthetic agar mouse-gray.
Streptomyces chrysomallus v.
fumigatus
II. Numerous spirals on aerial mycelium.
1. Aerial elycerol-glycine
agar grayish-rose. Practically no growth
mycelium on
on synthetic agar.
Streptomyces murinus
faciens, S.
2. Abundant growth on synthetic agar.
Aerial mycelium on
agar cream-colored.
glycerol-glycine
Streptomyces galbus v. achro-
mogenes
10. MAYAMA SYSTEM (1959)
Mayama (1959) concluded that morphol-
ogy, and types. of
growth on liquid media are the most impor-
serological reactions,
tant properties for the classification of
Streptomyces. On the basis of these prop-
erties, he divided the genus into seven
groups: S. olivaceus, S. lavendulae, S. aureo-
griseolus, S. albus, S. rimosus, and
S. reticult. In addition to these, he also listed
a number of species for which no group char-
acteristics were known, notably S. anti-
bioticus, S. fulvissimus, S. ruber, S. coelicolor,
etc.
Mayama (1959) and Mayama and Ta-
wara (1959) classified the genus Strepto-
myces into five sections and 14 series:
Section I. Aerial mycelium — irregularly
branched. Sporophores produced at the
GROUPS AND SPECIES OF GENUS STREPTOMYCES
FrGureE 30. Spore formation in Streptomyces species (Reproduced from: Shinobu, R. Mem. Osaka
Univ. Lib. Arts and Ed. B. Nat. Sei. 7, 1958).
terminal portion of the branching hyphae.
Series 1. Sporophores straight to flexu-
ous.
Series 2. Sporophores form open loops.
Series 3. Sporophores form spirals.
Section II. Aerial mycelium branches in tuft
formations. Sporophores produced at the
terminal portion of the branching hyphae.
Series 4. Sporophores straight to flexu-
ous.
Series 5. Sporophores form open loops.
Series 6.
Section ITI.
main stem. Sporophores produced at the
Sporophores form spirals.
Aerial mycelium forms long
terminal portion of side branches. Non-
verticillate.
Series 7. Sporophores straight to flexu-
ous.
Series 8. Sporophores form open loops.
Series 9. Sporophores form spirals.
Section IV. Aerial mycelium forms long
main stem. Sporophores produced at
terminal portion of side branches. Ver-
ticillate.
Series 10. Monoverticillate, straight to
flexuous.
Series 11.
Series 12.
Monoverticillate, spirals.
Biverticillate, straight to flexu-
OuS.
Series 15.
Section V.
Series 14.
Biverticillate, spirals.
No aerial mycelium.
11. NOMI SYSTEM
Finally a purely morphological system
may Nomi (1959) proposed a
division of the genus Streptomyces into eight
be listed.
morphological groups. He returned to an
earher concept of Drechsler (1919) that the
nature of the turn of the spirals, namely
sinistrorse and dextrorse, is an important
characteristic of Streptomyces species. He
recognized, however, that some cultures may
be rather indefinite in this respect.
A. Aerial hyphae somewhat flexuous or straight;
few long hyphae. The terminal filaments de-
velop into spiral-shaped sporophores.
1. Spirals sinistrorse.
104 THE ACTINOMYCETES, Vol. II
re
te ME SR
Y ob
10 11 12 13
oO
Puiate III. Morphological groups in the genus Streptomyces (Mayama, 1959) (For details, see text,
pp. 102-103).
Section I: series 1—3
Section IT: series 4—6
Section IIT: series 7—9
Section IV: series 10—18
B:
D.
ee
GROUPS AND SPECIES OF
a. Spirals long, extended to compact.
S. coelicolor, S. albogriseolus, S. fla-
veolus, S. parvullus
b. Spirals compact to compressed. None
found.
2. Spirals dextrorse.
a. Spirals long, extended to compact.
S. viridochromogenes
b. Spirals compact to compressed.
Streptomyces sp. No. 189
Most aerial hyphae long, straight or slightly
flexuous. They do not sporulate, but give rise
to short side branches whose terminal filaments
develop into spiral-shaped sporophores.
1. Spirals sinistrorse.
S. purpurascens
2. Spirals dextrorse.
Various unidentified forms.
. Aerial hyphae irregularly flexuous or wavy;
long hyphae absent. Terminal filaments form
spirals.
1. Spirals sinistrorse.
S. sulphureus
2. Curvature of spiral indefinite.
S. griseoluteus
Aerial hyphae long, straight, or wavy. They
give rise to short side branches, which develop
into spore-bearing hyphae containing spirals.
1. Spirals sinistrorse.
S. hygroscopicus, S. violaceoniger, S.
albus
. Aerial hyphae in clusters. The terminal fila-
ments develop into sporophores, both spiral
and nonspiral forming.
S. vinaceous, S. microflavus, S. fra-
diae, S. lavendulae, S.
roseochromogenes,
virginiae, S.
cinnamonensis, S.
S. phaeochromogenes
Aerial hyphae branch in clusters. No spirals or
loops.
S. venezuelae, S. tanashiensis, S.
bikiniensis, S. antibioticus, S. aureo-
faciens, S. olivaceus, S. nitrosporeus,
S. griseus, S. lipmanii, S. rutgersensis,
S. parvus, S. flavovirens, S. californi-
cus, S. vinaceus, S. ruber, S. caeruleus
Aerial hyphae long, straight or slightly flexu-
ous. Verticillate. No spirals.
S. reticuli, S. griseocarneus, S. echi-
mensis, S. hiroshimensis, S. salmoni-
cida, S. thioluteus, S. albireticuli, S.
netropsis
. Aerial hyphae somewhat flexuous or wavy.
Long hyphae and spirals are not produced.
S. albus (atypical), S. halstedii, S.
GENUS STREPTOMYCES 105
scabies, S. verne, S. griseolus, S.
erythreus (Plates IV and V)
12. OTHER SYSTEMS
Other systems have been proposed for the
the
Some of these systems are modifications or
supplementations of that presented in Ber-
gey’s Manual (7th ed., 1957), or modifica-
tions of one or the other of those outlined in
classification of genus Streptomyces.
this chapter.
One of these is the system outlined by
Routien (1959). The various species in-
cluded in the Streptomyces
divided into three major groups: (1) sapro-
phytes; (2) plant parasites or cultures
isolated from diseased plants or from soil in
which diseased plants were grown; (3) cul-
tures isolated from animal tissues. These
genus were
groups were subdivided on the basis of for-
mation and color of aerial mycelium (green,
brownish-purple to black, blue-gray or blue-
green, yellowish to orange, pink to rose, ete.).
The color of the substrate mycelium and the
various biochemical properties were then
used for further subdivisions. Morphology
(spiral formation, shape of spores) played
only a minor role in this system. See also
Sakai, 1959.
Summary of the Properties Used in
Subdividing the Genus
Streptomyces
Evaluation of the above systems of clas-
sification leads to the conclusion that sporo-
phore morphology has been given first or
second consideration by the great majority
of investigators. Lesser attention was paid
to the color of the aerial mycelium and the
nature of soluble pigments. Chromogenesis,
or pigment formation in protein media, Was
often given first position. Antibiotic pro-
duction and ecology received the least con-
sideration.
In Baldacci’s system of dividing the genus
into groups or series, the color of the sub-
THE ACTINOMYCETES, Vol. II
106
e ‘
a .
y A
ie a) ‘J
|
er ae
2x)
Q 2
Puate IV. Morphological types, according to Nomi (1959). a. represents a schematic presentation
of each type; b. gives the actual photograph.
A. Aerial hyphae flexuous or straight; spirals extended to compact (S. coelicolor).
B. Aerial hyphae straight; spirals on side branches (S. purpurascens).
(See continuation, next plate).
GROUPS AND SPECIES OF GENUS STREPTOMYCES LO7
PLate IV. (Continued)
C. Aerial hyphae wavy; spirals on terminal filaments (S. griseoluteus).
D. Aerial hyphae long, straight or wavy; sporophores as side branches; spirals produced (S. hygroscopi-
cus).
108 THE ACTINOMYCETES, Vol. II
ae
ine
a
oi
{
é
om
Ly.
-
fi
;
4
y
PLtate V. Morphological types, according to Nomi (1959). a. represents a schematic presentation of
each type; b. gives the actual photograph.
i. Aerial hyphae in clusters, terminal filaments developing into sporophores, both spiral- and nonspiral-
forming GS. lave ndulae, Se roseochromogenes ).
F. Aerial hyphae in clusters; no spirals or loops (S. antibioticus).
(See continuation, next plate).
GROUPS AND SPECIES OF GENUS STREPTOMYCES
ef,
lvl
30
Vv
50 20
| Nvvreluvistta quali
Prats V. (Continued)
G. Aerial hyphae verticillate; no spirals (S. hiroshimensis).
H. Aerial hyphae flexuous or wavy; no long hyphae and no spirals (S. albus).
109
L10
strate mycelium was taken as the basis for
the primary subdivision into sections, and
the color of the aerial mycelium for the
secondary subdivision into series. In the
systems used by Flaig and Kutzner (1954)
and by Ikutzner (1956), the color of the
aerial mycelium was used in connection with
that of the substrate mycelium. The system
proposed by Yamaguchi and Saburi (1955)
was based principally upon the structure of
the sporophores, the color of the aerial my-
celium being utilized in a secondary sub-
division; in the final characterization, ad-
vantage was taken of the production of
soluble pigments. A similar system was used
by Shinobu (1958). Krassilnikov (1941,
1949), Hesseltine ef al. (1954), Pridham e¢ al.
(1958), Etthnger et al. (1958), Mayama
(1959), and Nomi (1959) used morphological
criteria for the primary subdivision of the
genus.
Each one of the above systems has in it-
self certain serious limitations. It 1s neces-
sary, therefore, to combine several prop-
erties in order to bring out the characteristics
of the group or series, and especially those of
the species.
Proposed System of Classification of the Genus
Streptomyces into Groups or Series
In presenting the following system, full
cognizance is taken of the criticisms to be
directed against it, especially that the for-
mation of the melanin pigment is given
leading consideration, and that the pro-
duction of other soluble pigments as well as
of antibiotics is also given important con-
sideration. I have felt that because of my
own previous proposals, especially those in-
corporated in the various editions of Bergey’s
Manual, and my own interest in antibiotics,
the best I could do would be to modify this
system slightly. I hope that it will serve its
purpose in the future as it has done in the
past. The suggested series further broaden
my earlier concept of species-groups. In
THE ACTINOMYCETES, Vol. II
view of the fact, however, that it is desirable
for each group to be designated by a repre-
sentative species, it has been found neces-
sary, In some cases, to use a more recent,
well defined species rather than one used long
ago, for which no well established species is
now recognized. This is true, for example, of
the “‘chromogenes” series, which has been
designated as Phaeochromogenes, for which a
well recognized type culture is available.
I beheve that the system of classification
of the genus Streptomyces into series proposed
here is simple and convenient. The use of
ecological properties as a basis for the major
subdivision of the genus, as in the last edi-
tion of Bergey’s Manual, has been discarded.
The thermophilic forms have been, for the
part, transferred to other genera
(Chapter 11). The animal and plant isolates,
including both pathogens and saprophytes,
have been distributed throughout the genus,
most
among the various series, where they logi-
cally belong on the basis of their morphologi-
eal, cultural, and biochemical properties.
Both morphological (structure of sporo-
phores) and cultural (color of aerial myce-
lium, melanin formation) characters are
combined in the major subdivision of the
genus into subgenera and into series. Each
series is subdivided, on the basis of specific
cultural and biochemical properties, into
species. Formation of soluble pigments,
pigmentation, and antibiotic production are
also frequently taken advantage of in char-
acterizing species. To identify a new culture
properly, it is important to consider not only
the series subdivision and species classifi-
cation, but also the detailed description of
each organism. Before it can be decided
whether a newly isolated culture is different
from one already described, a study should
also be made of the varieties within the
species previously created, as well as possible
mutations and variations within the culture.
The names given for the various series are
GROUPS AND SPECIES OF GENUS STREPTOMYCES
the names of the type species within the
particular series. (See also Table 10.)
Genus Streptomyces Waksman and Hen-
rici, containing 16 series. Type species S.
albus (Rossi-Doria) Waksman and Henrici.
A. Subgenus Streptomyces Waksman, with
14 series. Type species Streptomyces (Strepto-
myces) albus (Rossi-Doria) Waksman and
Henrici.
B. Subgenus Streptoverticillium Baldacci,
with 2 series. Type species Streptomyces
(Streptoverticillium) reticuli (Waksman and
Curtis) Waksman.
A. Sporophores straight, wavy, or spiral-
forming. Subgenus Streptomyces
Subgroup A. MESOPHILIC
I. Melanin-negative
Series 1. Albus. This series is character-
ized by a white to light gray aerial myce-
lium, covering the whole of the substrate
growth; concentric rings may be formed. It
is melanin-negative. A faint brownish pig-
ment may be produced on organic media.
Sporophores are spiral-shaped, occasionally
broom-shaped. The species within this series
are usually strongly proteolytic, without
formation of bad-smelling products. It may
be argued that the type species S. albus is
no longer available and that many species
possess similar properties. This series and
this species must be recognized historically,
whatever the final the type
species to be adopted (Pridham and Lyons,
1960).
Series 2.
terized by a gray aerial mycelium, ranging
in color from light gray to mouse-gray or
smoke-gray to ash-gray to bluish-gray; it
may be white at first, later turning various
shades of gray. Substrate growth may be
decision of
Cinereus. This series is charac-
colorless or yellowish, turning gray to dark.
Frequently a soluble yellow pigment is pro-
duced. The sporophores are either straight
or spiral-shaped.
Nel
Series 3. Flavus. This series, as well, has a
long historical background; it was one of
the three groups so designated by Sanfelice
in 1904. It is characterized by a yellow or
vellow-orange to yellowish-brown substrate
growth, and by an aerial mycelium which is
white to yellowish to gray. A yellowish-
green to golden yellow soluble pigment. is
usually produced.
straight, or spiral-shaped.
Series 4. Ruber. This series is character-
Sporophores are long,
ized by a pink to orange to red substrate
growth, and by a white to yellowish to red
aerial mycelium. No soluble pigment is pro-
duced; occasionally a yellowish to brownish
pigment may be formed. Sporophores are
straight or spiral-shaped.
Series 5. Viridis. This series is character-
ized by a green to dark green substrate
growth, and by a white to gray to light
green aerial mycelium. Usually there is no
soluble pigment; occasionally a light green
pigment is formed. Sporophores are straight
or spiral-shaped.
Series 6. Violaceoruber. Substrate growth
is at first colorless, gradually becoming red
or blue; aerial mycelium is white to gray
with bluish tinge. The characteristic soluble
pigment is blue, frequently changing in color
with the reaction of the medium; it is blue
at an alkaline and red at an acid reaction.
Sporophores form spirals.
Series 7. Fradiae. This series is character-
ized by a yellow to orange substrate growth,
and by a powdery pink to seashell pink to
light orange aerial mycelium. Usually no
soluble pigment forms on synthetic or or-
ganic media; a pink pigment may occasion-
ally be produced. Sporophores are straight
or spiral-shaped. Species are strongly pro-
teolytic and antagonistic.
Series 8. Griseus. This series is character-
ized by colorless substrate growth, becom-
ing, in certain media, brown to almost olive-
black. Aerial mycelium is yellowish with a
greenish tint, or greenish-gray or sea-green.
112
No soluble pigment is produced. Sporophores
are straight or flexuous, producing tufts.
Series 9. Hygroscopicus. This series is char-
acterized by a colorless substrate growth,
which gradually becomes yellow, dark to
almost black. Aerial mycelium is white to
gray; it is often moist and even soft. Sporo-
phores are straight and spiral-shaped. No
soluble pigment is produced.
Il. Alelanin-positive
Series 10.
white to gray to buff. Substrate growth is
brown to black. Sporophores are straight
or spiral-shaped.
Series 11. Lavendulae. Aerial mycelium is
Scabies. Aerial mycelium is
lavender to rose or pink to vinaceous laven-
der. Substrate growth is colorless to cream-
colored. Sporophores are not flexuous, often
forming loops and loose or open spirals.
Series 12. Hrythrochromogenes. Aerial my-
celium is white with brownish shade. Sub-
strate growth is brown to black. Sporo-
phores produce spirals.
Series 13. Viridochromogenes. Aerial my-
celium is light green to olive-green. Sub-
strate growth is grayish-green to brown to
black. Sporophores produce spirals.
Subgroup B. THERMOPHILIC
Series 14. Thermophilus. This series com-
prises six species. These are listed in Chapter
1
B. Sporophores produce verticils.
Subgenus Streptoverticillium
AMelanin-negative
This and the
next series are largely characterized by the
Series 15. Cinnamomeus.
morphological structure of their sporulating
bodies. The sporophores produce verticils on
the primary or on the secondary branches of
the aerial mycelium, or on both. The spore
chains are straight or spiral-shaped. This
THE ACTINOMYCETES, Vol. II
group is further characterized by being
melanin-negative. The aerial mycelium is
white to pinkish to cinnamon-colored.
Melanin-positive
Series 16. Reticuli. This series is charac-
terized by the same morphological proper-
ties as Series 15, but it is melanin-positive.
The aerial mycelium is white to gray.
There is a considerable overlapping of the
different series. Frequently a given culture
may be placed in one series or another, de-
pending on the media and the conditions
used for growing the organism, not to
mention the idiosyncrasies of the observer.
Classification becomes particularly difficult
when one bears in mind the marked varia-
tions frequently observed between different
isolates of the same species, and the tendency
of individual cultures to mutate upon con-
tinued cultivation in artificial media. The
fact that identification is frequently based
upon comparison with published descrip-
tions rather than with type cultures has
resulted in the tendency to create new species
on the basis of minor differences, some of
which may be simple variations.
Most of the series are made up of non-
chromogenic forms (or those that produce no
melanoid pigments), although some of the
constituent species may produce faint brown
soluble pigments on certain media. Some of
these pigments result from lysis of the my-
celium of the organism; others may be quite
distinet and chemically different from the
typical melanoid or chromogenic pigments,
eg. the olive-green to olive-buff pigment
frequently produced by S. griseus. Fewer
series are composed of truly chromogenic
forms, those capable of producing brown to
dark brown or almost black soluble pig-
ments with protein-containing media.
The various ‘
quite distinct from those proposed by Bal-
dacci et al. (1954). They proposed, for ex-
‘series’? suggested here are
GROUPS AND SPECIES OF GENUS STREPTOMYCES 113
ample, a ‘‘Bostroem’’ series, for which no
true representative can be recognized at
present. Their series ‘‘Antibioticus’? and
“Caeruleus”’
reasons that need not be discussed further
here. Certainly, the idea expressed by Bal-
dacei et al. in 1955 that “it is not possible
to speak of a natural systemization of these
microorganisms at the present
knowledge... for the time being, one must
limit oneself to a classification aiming solely
at diagnosis and nomenclature,’ represents
a defeatist attitude. It is well illustrated by
his creation of a series named ‘‘Diastaticus.”’
state of
Here were included pigmented and nonpig-
mented organisms, chromogenic and non-
chromogenic, with such fantastic names as
A. rubrocyanodiastaticus, and such varieties
as atrodiastaticus. This is certainly a good
cause for confusion.
Similar criticism
grouping of the species proposed by Gause
et al. (1957). Whereas Baldacci used the
color of the substrate mycelium for primary
subdivisions of the genus into sections, and
the pigmentation of the aerial mycelium for
the further division of the into
series, Gause et al. (1957) omitted the sec-
tions altogether, and divided the genus
directly into series largely on the basis of
the pigmentation of the aerial mycelium.
Descriptions of 37 old and 71 new species
‘an be apphed to the
sections
were reported by a group of six collabora-
tors. The authorship was of a collective
nature, with all the possibilities for confusing
the credit to be assigned to each individual,
since it is stated that “the study of the
structure, classification, ecology and dis-
the
attention of large scientific collectives in a
tribution of actinomycetes occupies
number of institutes and universities.”
Proceeding from the fact that so many new
species have been recently created, in de-
scribing producers of antibiotics, these in-
vestigators assumed that this was further
‘cannot be accepted for other
proof that the old systems of classification
were insufficient. Although it was recog-
nized that the pigmentation of the aerial
mycelium, the major criterion for classifica-
tion purposes, could change on continued
incubation, as in the case of their Group I,
“cultures with lavender and brownish-rose
pigment may change in color to salmon, red,
and pale terra cotta,’ nevertheless, 15 series
were adopted. This fact alone would tend to
cast doubt upon the significance of recog-
nizing major groups solely on the basis of
pigmentation of the aerial mycelium. In
establishing the species, structure of the
sporophores was used in some cases; in
others, the pigmentation of a single medium,
frequently unknown in composition, was
used.* To complicate the situation further,
authors of old species and emendations of
species were incorrectly credited, providing
a potential source of confused nomenclature.
Lieske (1921) was the classical ‘‘tumper,”’
largely because of the limitations imposed
by the use of complex organic media, and
because he was not aware of some of the
characteristic morphological and cultural
properties of the organisms, brought out
particularly on synthetic media. The “‘split-
ting” attitudes of Baldacci, Gause,
certain others have brought the system of
classifying this important group of organ-
4 and
isms to undesirable extremes.
In the decision to classify the genus Strep-
tomyces into 16 series, it is well understood
that in time other series will be added; some
of those presented here may eventually be
spht into two or more series; some of the
varieties may be raised to the status of
species; or some of the species may be raised
to the status of series.
The problem of whether antibiotic produc-
tion is a species characteristic is still un-
settled. Undoubtedly, the
different antibiotics can be combined with
production of
* Hottinger’s, to which no reference is given.
114 THE ACTINOMYCETES, Vol. II
certain other distinct properties, such as
pigmentation, morphology, and carbon utili-
zation, to justify the creation of new species.
This has actually been done for the separa-
tion of S. griseinus from S. griseus and the
raising of the latter to a series status. In
other cases, however, the mere formation of
a different antibiotic without other accom-
panying differences hardly justifies, for the
present at least, the creation of new species.
Chapter 6
Series and Species of the Genus
Streptomyces
The genus Streptomyces was created in
1943, to separate certain aerial mycelium-
producing actinomycetes from the rest of
the order Actinomycetales. Although there
is considerable overlapping between species
placed in this genus and those of Nocardia
and some of the thermophilic groups, there
are certain important properties that may
said to characterize this genus, thus
separating it, if not for any other reason
than that of convenience, from the others.
The major important characteristic prop-
erties that distinguish the genus Streptomy-
ces from the others can be briefly summar-
ized as follows:
be
1. A more or less branched, nonseptate,
substrate or vegetative mycelium (stroma)
is produced.
2. Growth takes place either on the sur-
face of agar or gelatin media or penetrates
deep into the medium, forming a compact,
often leathery mass, designated as a colony.
During growth in stationary liquid
media, no turbidity is produced except on
9
oO.
lysis; the masses of growth appear as clumps
or compact Masses.
4. The surface colony gradually becomes
covered with an aerial mycelium, though
this occasionally may not occur.
5. The aerial mycelium produces sporo-
genous hyphae or fruiting bodies, which are
straight, or in the form of tufts, or curved,
spiral-shaped, or verticillate.
ibs)
6. The sporophores carry chains of single-
celled spores (or conidia), which vary in
shape from spherical to oblong or cylindrical,
and also in surface appearance when viewed
with the electron microscope.
7. The vegetative growth, the aerial my-
celium, and the spores en masse frequently
are colored in a characteristic manner; the
color may also dissolve into the medium,
producing a
‘
‘soluble pigment.”’
8. The species are aerobic and meso-
philic, nonacid-fast and gram-positive.
The genus Streptomyces comprises, by far,
the largest number of species of actinomy-
cetes now known to occur in nature. The
various species belonging to this genus differ
in cultural,
physiological, and biochemical properties.
greatly their morphological,
They include the majority of antibiotic-
producing actinomycetes. The growing eco-
nomic importance of these organisms has
tended to increase the need for the separa-
tion of the genus into groups, each of which
would contain one or more species. This
need has recently been further emphasized
by the creation of numerous additional spe-
cles.
The color of the aerial and substrate my-
celium, the morphology of sporophores, and
the formation of melanin pigments have
been largely used for the separation of the
genus Streptomyces into series and species.
lor the supplementary characterization of
116 THE ACTINOMYCETES, Vol. II
the species, the formation of nonmelanin
pigments, the ecology of the organisms, and
some of the biochemical properties (notably
antibiotic formation) have been utilized.
This is also true of their practical utiliza-
tion for the production of enzymes, vita-
mins, or antibiotics.
It would appear that morphological prop-
erties might offer a natural and stable basis
for a system of classification of these organ-
isms. Unfortunately, certain characteristic
morphological features of the genus Strepto-
myces undergo variation, depending upon
the nutrition of the organisms and upon the
environment. This tended to suggest, at
first, the inadvisability of considering mor-
phology as the major basis for the classifica-
tion of the genus Streptomyces. This was true,
for example, of Drechsler’s idea of consider-
ing the type of curvature of the spiral-form-
ing aerial hyphae as a basis for classification.
It was also true of Waksman’s suggestion
that the mode of branching of the sporo-
phores might be used for this purpose. The
ideas of Krassilnikov (1941, 1949) in empha-
sizing the size and shape of the spore would
also meet with similar criticism. Flaig et al.
(1955), as well as Ettlinger et al. (1958), pro-
posed use of the nature of the spore surface
as a species characteristic; unfortunately,
this property, depending as it does upon the
use of the electron microscope, has not been
readily enough established to enable the sep-
aration of the genus into groups and species.
Certain morphological properties are
now, however, well recognized and can be
utilized for the separation of certain groups
of organisms belonging to the genus Strepto-
myces. Such groups possess sufficiently well
defined morphological features to differen-
tiate them from the rest of the genus. This
is true particularly of those forms that
produce radiating sporulating hyphae (verti-
cils), with straight or spiral-shaped branches
on the main sporophores or on the side
branches. This property makes it possible
to distinguish these particular forms from
the majority of other species of Streptomy-
ces, Which produce either straight, flexuous,
curved, or spiral-forming sporophores. Sev-
eral systems of classification of the genus
Streptomyces into series (Hesseltine ef al.,
1954; Shinobu, 1958b) took full advantage
of the verticil-producing property; Baldacci
(1959) went so far as to suggest placing the
latter into a separate genus. The separation
of the spiral-forming from the straight
sporophore-producing types into separate
groups has also been frequently suggested.
In view of the above limitations, the only
conclusion that can be reached is that, for
the present at least, a logical system of
separation of the genus Streptomyces into a
number of distinct series should be based
upon a combination of several of the mor-
phological and physiological properties. It
is proposed here to divide the genus into
16 series. This system, likewise, is open to
criticism: (a) there is left, for example, con-
siderable room for a certain amount of over-
lapping in some of the major properties which
characterize the various series; (b) the posi-
tion of a species within a series is not always
well defined, and some of the species could
frequently be placed with as much Justifica-
tion in one series as in another; (c) there is,
further, a lack of uniformity in characteriz-
ing the various series: In some instances
color of the aerial mycelium or of the sub-
strate growth is used, and in others the
formation of soluble pigments is emphasized.
Fully recognizing the above limitations,
however, I feel that a sound basis has been
laid, taking full advantage of the knowledge
now available, for dividing the genus Strepto-
myces into series. As further information
accumulates, the system can easily be modi-
fied, since it lends itself readily to various
changes and modifications.
A detailed characterization of the various
series 1s presented here. Some of the series
are described in greater detail than others.
SERIES
This is due either to their longer historical
background or to their capacity to form
important economic products, especially an-
tibiotics.
In characterizing each series, the follow-
ing properties have been given special con-
sideration.
a. Morphological properties, — notably
structure of sporophores and spores.
b. Color of aerial mycelium on synthetic
media.
ce. Formation of soluble brown pigment
(melanin) on protein media.
d. Spore surface.
e. Other characteristic properties, such
as color of substrate growth, formation of
soluble, nonmelanoid
pigments, rate of
proteolysis, or production of specific anti-
bioties.
A summary of the properties of the 15
series within the genus Streptomyces is given
n Table 10.
AND SPECIES OF
GENUS STREPTOMYCES 117
I. Series Albus
Characteristic Properties
a. Sporophores produce spirals; spores
spherical to oval.
b. Color of aerial mycelium white.
c. Melanin-negative.
d. No soluble pigment produced (except
a faint brown pigment on certain media).
e. Weakly proteolytic and weakly antago-
nistic.
The Albus series comprises a large num-
ber of organisms, characterized by the pro-
duction of
substrate
a typical leathery and compact
growth, various
media. Aerial mycelium is snow-white to
colorless on
white in color, assuming various shades as
the culture grows older. The sporophores
are long and form spirals; the spores are
spherical to ovoid. The various strains grow
well on both organic and synthetic media.
They vary greatly in their proteolytic and
diastatic properties. As a rule, this group of
TABLE 10
Characteristic properties of various series of Streptomyces
SA Name of series Mee Aerial mycelium Color of growth forma Type species
1 Albus — White Colorless + .§. albus
2 Cinereus = White to gray Colorless to yel- | +— | S. craterifer
low
3 Flavus — Mouse-gray Yellow + | 8S. flavus
4 Ruber — Rose Red +— | §. ruber
5 Viridis —_ Gray to green Green +— | S. viridis
6 Violaceoruber - Gray Red to blue + | S. violaceoruber
7 Fradiae - Pink to rose Yellow to orange +— | S. fradiae
8 Griseus — Grass-green Colorless to olive- — |S. griseus
buff
9 Hygroscopicus — White to gray Dark gray to black + |S. hygroscopicus
10 | Scabies + Gray Brown to black + |S. scabies
11 Lavendulae a Lavender Colorless +— §. lavendulae
12 Erythrochromo- + Yellowish Orange +— |S. erythrochromo-
genes genes
13 Viridochromo- + Green to olive- | Brownish to green + | S. viridochromo-
genes green genes
14 Cinnamomeus — Pinkish Yellowish — SS. cinnamomeus
15. | Reticuli + | White to gray Colorless +— | S. reticulr
118
organisms, so far, has not been reported as
containing any significant antibiotic-pro-
ducing forms. Although one of the first
preparations possessing certain antibacterial
properties ever recorded for a culture of an
actinomycete was said to have been obtained
from a member of the Albus group (Gratia
and Dath, 1925), it is open to question
whether the particular culture was a true
S. albus. According to Pridham and Lyons
(1960), this organism should be considered
as more closely related to the Griseus group.
Species belonging to the Albus series are
found in soil and in dust. Various early in-
vestigators, notably Almquist, Gasperini,
Rossi-Doria, Beijerinck, and Sanfelice, re-
ported the isolation of organisms belonging
to this series.
Various systems of classification of the
Albus series have been proposed. Attention
may be directed here to the fact that at one
time or another all the sporulating actino-
mycetes, especially the saprophytic forms,
mostly now recognized as belonging to the
venus Streptomyces, were classified (see Bei-
jerinck, 1900, for example) ito two groups:
(1) A. albus (Streptothrix alba), comprising
those forms that produce a white aerial
mycelium and no soluble pigment; (2) A.
chromogenus (Streptothrix chromogena), i-
cluding those forms that produce a black
pigment on protein media. Duché appeared
to follow this system as late as 1934, since
he included in his monograph (Duché,
1934) on the actinomycetes only those spe-
cies that were said to belong to the A. albus
group. In view of the significance of the
specific name ‘‘albus,” representing the type
eulture of the genus Streptomyces, it may be
of interest to trace the usage of this name in
the literature on the actinomycetes.
In presenting this historical summary, the
writer has taken full advantage of the com-
ments this group made by
Baldacci (1939), who is frequently quoted
Baldacei
concerning
here almost verbatim. recorded
THE ACTINOMYCETES, Vol. II
about 30 some of which are
listed in Table 11. (Others were not included,
since they are not considered as typical of
the group).
The name ‘‘alba”’ was first applied to an
actinomycete culture by Rossi-Doria (1891).
He established the characteristics of this
organism, indicating its synonymy with the
cultures previously characterized by Alm-
quist (1890); he also identified it with a
culture designated as Streptothrix Foerster,
isolated from the air by Gasperini (1890).
Xossi-Doria refused to accept the identi-
fication of this organism with Streptothrix
Foersteri Cohn. He said: ‘Nothing in
the description given by Cohn can justify
such an idea. In that description, in fact,
only generic characters are given; of spe-
synonyms,
cific characters there does not exist even a
shadow.’ The three cultures of Almquist
appeared to differ little among themselves.
Since one (culture I) was said to form a
white crust changing in time to gray, Bal-
dacci preferred to exclude it from the syno-
nymity with the A. albus. Gasperini (1894)
recognized the difference between A. albus
and A. chromogenus. The latter possessed
chromogenic properties, the pigment diffus-
ing into the substrate.
Santelice (1904) was the first to divide
into three groups the actinomycetes now
recognized as belonging to the genus Strepto-
myces, using S. albus as the representative
of the first of these groups. He noted that
some of the cultures belonging to this group
may produce a black pigment when grown
on potato. He added quite significantly:
“On the basis of this observation, a super-
ficial observer may create a new species out
of a pigmented culture without considering
the fact that it originated from Str. alba.”
Krainsky (1914) isolated from garden soil
a culture which he described as A. albus.
This culture produced a well developed
growth, white at first, then becoming gray
on certain media such as glucose agar and
SERIES AND SPECIES OF GENUS STREPTOMYCES
TABLE 11
Nature and dimen-
pcos ene Author sions of spore
“
Cladothrix dichotoma | Macé a E
Streptothrix Foersteri | Gasperini Oval (1.0-1.5)
Streptothrix n. 2 Almquist +
Streptothrix n. 3 Almquist +
Streptothrix alba Rossi- Doria | +
Actinomyces albus Krainsky Oval (1.0)
A, albus Waksman and
Curtis by 1.1-1.4)
A. albus | Jensen Rectangular (0.4-0.5
by 2.4)
A. albus Duché +
A. chromogenus Gasperini Oval (1.5)
Cladothriz odortfera Rullmann + (1.0)
A. thermophilus Berestnew +
A. thermophilus Gilbert + (0.5-0.6)
A. thermodiastaticus | Bergey Oval
A. sanninii Ciferri Round
A. almquisti | Duché | +
A. gougeroti Duché a
Streptothriz gedanen- | Scheele and oo
a | |
sis I Petruschky
Streptothrix candida | Petruschky +
Streptothriz lathridii | Petruschky a
Cladothrix invul- | Acosta and G. ao
nerabilis Rossi
Spherical, oval (1.2-1.6
Comparative characters of Streptomyces albus and related species (Baldacci, 1939)
Mycelium
119
Proteolytic
action
Chrom-
bes ogenesis
Substrate Aerial Gelatin | Milk
White a
Colorless to yellow-| White _ +
ish
White ~ a
White - ~
Colorless to black | White ~ _
Colorless White (less often = +
gray)
White or gray Cream or gray — + a
Cream or yellow- | White >
ocher
White-yellow White = ~~
| Ocher to black White +
| White or chalk | Weak | +
white
| White
Gray-yellow Gray ar SF
Colorless White ar =
Colorless White or ivory oF Weak
white
Yellowish - ~
Greenish | White } —
Colorless | White Weak
Colorless | White 3 te
Colorless White + +
| White - +
gelatin. The aerial mycelium was produced
readily, the medium remaining colorless.
The spores were oval, 1 yw in size. Gela-
tin was liquefied. Nitrate was reduced. The
culture had no diastatic action on starch.
Waksman and Curtis (1916) isolated from
soil cultures of an organism considered to
be A. albus. It was similar to that of Kvain-
sky, although the exact identity of the two
was doubted. The diagnosis of Krainsky
was, therefore, amended. The aerial my-
celium appeared either white or gray, ac-
cording to the composition of the medium;
the substrate growth varied from white to
gray. The sporophores produced short and
rare spirals. The spores were 1.2 to 1.6 by
1.1 to 1.4 uw. The culture was nonchromo-
genic, hydrolyzed starch, reduced nitrate,
and liquefied gelatin. Jensen (1931) also
isolated a culture of A. albus from soil. The
aerial mycelium was constantly white, and
the substrate growth cream-colored
yellow-ochre. The culture was said to pro-
duce cylindrical spores, 0.4 to 0.5 by 2.4 wu.
Duché (1934) created excessively
broad “A. albus group.’ He was severely
criticized by Baldacci (1939), who said that
‘df it is meant by ‘albus group’ those species
or
an
that produce white aerial mycelium in cul-
ture, they are many more in numbers than
those described by Duché. These are also
the ‘viridis’ species and those that show
analogy with the ‘flavus’ forms.’? Baldacci
further emphasized that A. albus is a well
characterized species that does not permit
with the ‘‘albus
group”? of Duché comprised 18 species, of
confusion others, while
which 15 were new ones; among these were
120 THE ACTINOMYCETES, Vol. II
such forms as A. viridis, A. albidoflavus, and
A. alboflavus, which definitely belong to
other groups.
In his morphological study of the actino-
mycetes, Duché recognized five types of
sporulation, none of which was used for
systematic purposes. The relationship of
aerial hyphae, spirals, and spores was not
sufficiently emphasized. Duché stated that
colonies may also originate from arthro-
spores, implying thereby that any mycelial
fragments will reproduce and multiply in
the culture. He spoke, however, of ‘“‘mono-
sporic”’ although he used the
method of successive dilutions and not that
of single-spore isolation. He documented
the various interpretations of the species
A. albus, stating at first that the description
of Waksman “‘ne correspond pas tout a fait
aux type albus de Gasperini, Rossi-Doria
et Krainsky.”’ On comparing his own cul-
ture with the preceding ones, he stated,
“T’espece de Waksman and Curtis semble
posséder toutes les propriétés de celle de
Krainsky.... Notre espece resemble aux
deux précédentes.”’ Baldacci concluded that
the work of Duché, after trying to prove the
diversity of the various interpretations in
the literature, had not attained its purpose
of establishing what the species A.
colonies,
albus
should be.
In proposing the genus Streptomyces in
1943, Waksman and Henrici stated: ‘‘We
have selected as the type species of this
newly named genus, Streptomyces albus
(Rossi-Doria emend. Krainsky) comb. nov.
This species was formerly known as Actino-
myces albus WKrainsky and first described as
Streptothrix alba Rossi-Doria. This is one
of the commonest and best known species
of the group, and, although it may later be
subdivided into further species, it 1s at
present as definite as any other. It has been
recently studied intensively by Duché (1954)
and by Baldacci (1939). It is colorless, with
ovoidal
white aerial mycelium, forming
spores in coiled chains on lateral branches
of the aerial hyphae. It is proteolytic, lhque-
fying gelatin and peptonizing milk with the
production of an alkaline reaction in the
latter. It does not produce any soluble pig-
ment either on an organic or synthetic me-
dium, but does produce a characteristic
earthy or musty odor.”
Pridham and Lyons (1960) have recently
made a comprehensive analysis of the pres-
ent status of Streptomyces albus. Their study
was based upon a detailed examination of
55 cultures collected from various sources.
They came to the conclusion that ‘‘there
has existed since about 1916, two entirely
different concepts with regard to the nature
of Actinomyces (Streptomyces) albus. One
concept centers around strains with the
following characteristics: flexuous fruiting
bodies, colors of aerial mycelium in tints
and shades of olive-buff (yellowish-gray or
tan); nonchromogenicity (nability to form
brown, deep brown, or black diffusible pig-
ments in organic substrates); and marked
abundance in nature” (these strains are now
considered as comprising members of the
Griseus group). “The other concept con-
cerns strains that are characterized by coiled
or spiralled fruiting bodies with catenulate
ovoldal spores; by aerial mycelium colors
generally interpreted as cretaceus (chalk-
white, often with faint tinges of pink); by
nonchromogenicity (inability to form brown,
deep brown, or black diffusible pigments in
organic substrata); and by their relative
rareness in nature” (these strains are now
considered as Albus group proper).
Morphologic Characters
Various methods were used in the study
of the morphology of S. albus. Baldacci
observed two types of mycelium. One was
hyaline, not less than | uw in diameter, rami-
fying more or less abundantly, and having
an undulated appearance. The ramification
starts perpendicularly from the point of
SERIES AND SPECIES OF GENUS STREPTOMYCES 12]
intersection, but it can follow in other di-
rections or assume a Wavy appearance. This
mycelium originates directly from the germi-
nating spores and can be rather abundant.
It corresponds to the first vegetative growth
and is designated as ‘“‘substrate mycelium.”
The second type of mycelium is more dis-
tinctly visible than the first. It is larger in
diameter (1.1 to 1.4 yw); it is subhyaline
with a tendency to assume yellowish colora-
tion. This mycelium carries abundant sporog-
enous hyphae, scarcely ramified. The ex-
tremities curve to hook shapes and succes-
sively turn to spirals. This mycelium is
white and is superimposed on the substrate
growth; in time, it turns to dirty white or
milky white, powdery or crusty. It is desig-
nated as ‘‘aerial mycelium.”
The branches of the aerial mycelium be-
come sporophores and give rise to spores
that are formed by contraction. The spores
are short, oval in shape, white, and not
without a certain polymorphism, appearing
sometimes as short rods. One may also ob-
serve round forms, but they must be inter-
preted as spores seen in a vertical projection.
The spore dimensions are 0.6 to 0.7 by 1.2
u. According to Baldacci, the spores are
smaller than those observed by Jensen and
longer than those described by Drechsler.
Baldacci was not sure, however, that Drech-
sler’s culture corresponded to S. albus. The
difficulty in reaching an agreement con-
cerning the shape and measurement of the
spores is not due, as claimed by Duché, to
their small size, but to the time at which the
measurements are taken and observations
made. When the spores are united in chains
in the sporophores, they appear longer and
rectangular; if measured when they are
spread in the preparation, they appear
shorter. It is natural, therefore, that only
the shape and dimension in the latter case
be accepted. It should be noted that the
free spores are not distributed on the slide
in a uniform manner, because many remain
attached to the glass at their smaller sur-
face, and thus appear round. Spirals are
abundant.
The degeneration of the hyphae can be
observed in the old substrate mycelium:
protoplasm
zones of with empty
simulating arthrospores can be seen. This
spaces
phenomenon led earlier investigators, includ-
ing Gasperini, to make unjustified generali-
zations. This is also evident in the claims of
Lachner-Sandoval, Vuillemin, and Grigora-
kis, who observed this false sporulation. Al-
though these authors did not always specify
the particular species used in their studies,
Baldacci was inclined to think that they
had to do rather with a Nocardia; the vege-
tative mycelium of the latter, when its
growth is arrested, subdivides into frag-
ments that look like bacillary elements.
This type of fragmentation was studied by
Mrskov and by Jensen. The ‘
irregulars”’ of Duché and others bring out
very clearly this type of degeneration, which
was erroneously interpreted as a type of
sporulation.
‘arthrospores
Cultural Characters
S. albus produces a colony in the form of
a growth adherent to the substrate; it is
wrinkled and colorless. The aerial mycelium
appears first in the drier portion of the
colony; it is chalk-white in color, as if lime
had been sprayed over it. On aging, it turns
to ivory-white; on some media, such as
nutrient assumes a grayish tint.
With age, growth of the culture becomes
agar, it
opaque or even yellowish, comparable to
the color of the substrate, as can be ob-
served in the cultures where the aerial my-
celium is not produced. The formation of
the aerial mycelium appears to correspond
to the presence of water of condensation,
the aeration of the cultures, and various
other factors, independent of the strain or
variety. The aerial mycelium is more or less
abundant and may cover the entire colony.
122 THE ACTINOMYCETES, Vol. II
Concentric ring formation may be observed
in smaller colonies, as described by Rossi-
Doria. The cultures produce the character-
istic moldy or soil odor.
On synthetic agar, the initial develop-
ment of the culture is characterized by a
dusty or powdery white, dry growth, form-
ing furrows or concentric rings. The sub-
strate mycelium is formed in a thin, scarcely
visible layer over the agar. The colony does
not assume a vigorous aspect. The white
aerial mycelium appears late. Pigmentation
of the agar is seldom observed, except for
a few strains that show feeble chromogenesis
in this medium.
On potato, the development is rapid, with
small, partly confluent colonies or in the
form of extended membranous growth that
becomes covered with white aerial myce-
lium.
Biochemical Properties
Temperature: Optimum 24-28° (24—
44°)C,
Gelatin: Liquefied.
Starch: Diastatic action variable.
Sucrose: Inverted.
Nitrate: Reduced to nitrite.
Antagonistic properties: The organisms
belonging to the S. albus group are usually
weak antagonists. Some cultures possess
activity against gram-positive bacteria.
Species
KrassilInikov (1949) included 19 species
in the Albus series. He used a combination
of different criteria for their separation and
identification. Tor separation of the cul-
tures, he considered the odor produced as
the major criterion, which is rather unrelia-
ble. Use of this criterion is largely responsi-
ble for the inclusion, in this group of species,
of forms designated as aromaticus, odoratus,
odorifer, putrificus, ete. IKrassilnikov also
considered, for identification purposes, the
shades of white in the aerial mycelium, tem-
flocculus, A.
perature relation, proteolytic and antago-
nistic properties, the secretion of a brown
substance, growth in acid media, production
of ammonia and H.S, decomposition of
rubber, and formation of coremia. He in-
cluded in this group various thermophilic
and thermotolerant organisms.
Gause et al. (1957) divided the Albus
series, on the basis of the color produced on
a complex organic medium, into three sub-
groups, comprising five species and one
variety:
a. Medium not pigmented: A. candidus,
A. candidus var. alboroseus, and A.
albidoflavus.
b. Medium colored brown: A. longisporus
and A. mirabilis.
c. Medium
rubidus.
The above characterization fails to recog-
nize some of the fundamental cultural prop-
erties of actinomycetes, namely, the produc-
tion of melanin pigments in protein media
and the structure of the sporophores. The
resulting subdivision of such a group into
albo-
colored brownish: A.
subgroups thus loses all significance. Only
one of the above species (A. albidoflavus) is
found in’ Krassilnikov’s ‘‘albus’’ — series.
Kutzner (1956) reported that he had ob-
tained four strains of S. albus from different
institutions and found them to be identical.
Baldacci placed in the Albus series the
following organisms, either because they
were considered as synonyms or because
they were believed to be closely related:
Actinomyces albus, A. acidophilus, A. alm-
beddardi, A. chromogenus, A.
A. exfoliatus, A. farcinicus, A.
gedanensis, A. gelaticus, A.
gougeroti, A. heimii, A. kimbert, A. liesket,
A. listeriit, A. malenconi, A. reticult, A. soma-
liensis, A. sanninii, A. saprophyticus, A.
thermophilus, A. wpcotti1, A. willmorec; Clad-
othrix dichotoma, C. liquefaciens, C. imvul-
nerabilis, C. odorifera; Oospora doriae, O.
alpha; Streptothrix alba, Str. candida, Str.
quisti, A.
erythreus,
SERIES AND SPECIES OF GENUS STREPTOMYCES 12
dassonvillei, Str. foerstert, Str. graminearum,
Str. leucea, Str. lathridiz, and Str. pyogenes.
Such a large conglomeration defeats com-
pletely the purpose of grouping, since the
above forms vary greatly morphologically,
culturally, and physiologically.
A number of species can, however, be in-
cluded in this series. It is sufficient to men-
tion S. albus, S. calvus, and S. niveus.
Il. Series Cinereus
Characteristic Properties
a. Sporophores straight or spiral-shaped.
b. Color of aerial mycelium white to
gray; occasionally dark humid stains or
guttation drops.
ce. Growth usually colorless, occasionally
yellow to tan.
d. Melanin-negative.
When
aerial mycelium, the color is characteristi-
‘ally gray. Although it may be white at
first, it changes to various shades of gray,
members of this series form an
ranging from light gray to mouse-gray to
bluish-gray or even vinaceous-gray or black-
ish-gray. Frequently white spots are pro-
duced in the aerial mycelium. The substrate
growth is often colorless or gray, occasion-
ally becoming yellowish to buff-colored; it
is either opaque or somewhat slimy; the
reverse 1s usually colorless, occasionally
turning yellow to tan. It is melanin-nega-
tive. Occasionally a yellowish or brownish
soluble pigment may be produced. The
sporophores are straight, often formed in
clusters or tufts; they may also produce
spirals that are either open or compact.
This series is widely distributed in nature.
It comprises, in addition to well described
species given here, a great number of incom-
pletely described forms, as shown in Chapter
i:
A variety of antibiotics have been found
to be produced by members of this series.
Oo
Some Characteristic Species
The following species may be tentatively
included in this series: S. craterifer, S. inter-
medius, S. parvullus, and S. cellulosae.
One may also include in this series various
organisms described by Krassilnikov (1949),
including his emendation of <A. griseus
Kkrainsky (not S. griseus Waksman), A.
variabilis, and A.
This is also true of some of the forms de-
by Gause et al. (1957) under such
griseus griseus zonatus.
scribed
names as A. rubiginosus, A. griseomycini,
A. werint, A. acrimycint, A. acrimycini var.
atroolivaceus, and A.
globosus, A. griseoru-
bens.
IIL. Series Flavus
Characteristic Properties
a. Sporophores long, spiral-shaped; spores
spherical to oval.
b. Aerial mycelium white to gray to
mouse-gray ; color of growth yellow to golden
yellow.
c. Melanin-negative.
d. Yellowish-green to golden soluble pig-
ment may be excreted into the medium.
e. Strongly proteolytic and antagonistic.
The Flavus series includes a large number
of organisms, widely distributed in nature.
The members of the series vary greatly in
some of their cultural, biochemical, and
morphological properties when grown on
artificial media. This group has been recog-
nized 1891,
scribed a culture under the name of Strepto-
thrix albido-flava. Another similar culture
since when Rossi-Doria de-
was soon described by Gasperini (1892) as
Actinomyces albido-flavus. Sanfelice (1904)
designated the second of his three groups as
Str. flava, comprising organisms isolated
from the air. Caminiti (1907) was inclined
to include in this group various pigmented
forms, such as Str. cztrea and Str. chromo-
gena.
Numerous organisms belonging to the
124
Flavus series have been isolated from soil,
dust, and other natural substrates by Krain-
sky, Waksman and Curtis, and others. They
have been designated by a variety of names,
such as S. alboflavus, S. aureus, S. citreus,
S. griseoflavus, and S. flaveolus.
The Flavus series is characterized by
cream-colored to yellow or golden yellow
growth on most artificial media. The aerial
mycelium is usually white to gray to mouse-
gray. The sporophores are long, usually
spiral-shaped. The spores are spherical, us-
ually 0.7 » in diameter. No brown pigment
is produced on protein media. A yellowish-
green to golden pigment is often formed in
synthetic and organic media. The various
species in this group are strongly proteolytic
and diastatic. Sucrose is inverted. Nitrate
is reduced. Many of the strains are strongly
antagonistic and are able to form active
antibiotics, some of which have found ex-
tensive application as chemotherapeutic
agents.
KrassilInikov (1949) recognized 13 distinct
species as belonging to the Flavus series.
Baldacci (1939), however, subdivided the
actinomycetes with the characteristics of
the Flavus series (various species producing
vellow or golden growth) into a number of
series: “aureus,” “albidoflavus,” “sulphur-
eus,”’ “antibioticus,’ and ‘‘hygroscopicus.”’
Baldacci et al. (1954) included in the ‘‘Aw-
reus”’ series such species as S. aureus, S.
aureofaciens, S. citreus, S. fimicarius, 8.
flavus, S. flaveolus, S. fordit, S. griseoflavus,
S. hygroscopicus, S. microflavus, and S. par-
DUS.
A culture of an organism isolated by
Takahashi (1953) in Japan was identified
by him as S. flaveolus Waksman. To validate
this identification, his description of this
culture is presented in Table 12 alongside
Waksman’s description of the original type
culture. These data show that, in spite of
minor variations in color characterization,
quantitative differences in gelatin lquetfac-
THE ACTINOMYCETES, Vol. II
tion and nitrate reduction, and even in
differences in antibiotic production, the
identification of the species appears to be
correct.
The same is true of the characterization of
S. parvus. The original culture of this organ-
ism, which was used as the basis for its
description in Bergey’s Manual, has died out
in the collection. Benedict, of the Northern
Regional Research Laboratory of the U.S.
Department of Agriculture, isolated from a
sample of soil collected in West Africa a
culture which he identified as S. parvus. A
comparison was made of the culture origi-
nally isolated and described by Krainsky
(1914), the culture isolated by Waksman
and Curtis (1916) and reported in Bergey’s
Manual, and the new culture of Benedict
(Table 13). The results point definitely to
the identity of the three cultures, thus prov-
ing again that accurate identification of
some species can be made by comparing
freshly isolated cultures with written de-
scriptions of type isolates.
Finally, a comparison was made (Waks-
man, 1957) of two published descriptions of
S. flavus and S. griseoflavus, together with
recent descriptions of two cultures that
have been raised to the status of new species,
namely, S. aureofaciens and S. rimosus, both
important producers of antibiotics. The re-
sults, presented in Table 14, show that S.
aureofaciens and S. rimosus are sufficiently
different from S. flavus and S. griseoflavus
to warrant the creation of new species. S.
aureofaciens is characterized by a deep gray
aerial mycelium, by a lack of or limited
spiral formation, by lmited proteolytic ac-
tivity upon gelatin and milk, and by poor
growth on nutrient agar. It was concluded
that properties differentiated this
culture sharply from the two older cultures.
these
S. rimosus is characterized by poor growth
on synthetic agar and by the formation of
abundant spirals in its aerial mycelium.
together with certain
These properties,
SERIES AND SPECIES OF GENUS STREPTOMYCES
—_
~
oO
TABLE 12
Identification of Streptomyces flaveolus (Waksman, 1919; Takahashi, 1953; Waksman, 1957)
Characteristics
Morphology
Sporophores
Spores
Synthetic agar
Substrate growth
Aerial mycelium
Soluble pigment
Calcium
NH,Cl agar
Substrate growth
Aerial mycelium
Soluble pigment
Nutrient agar
Substrate growth
Aerial mycelium
Soluble pigment
Gelatin
Substrate growth
Aerial mycelium
Soluble pigment
Liquefaction
Potato
Substrate growth
Aerial mycelium
Color of plug
Glucose broth
Substrate growth
Aerial mycelium
Soluble pigment
Milk
Nitrate reduction
malate- |
| Cream-colored
Waksman Takahashi
Numerous spirals on all media
Oval to elliptical
Numerous spirals on synthetic media
Spherical or oval, 0.8 by 1.2 u
Light sulfur-yellow turning cad-
mium-yellow
White with ash-gray patches
Empire-yellow
Antimony-yellow to chamois-colored
White, later smoke-gray
Buff-yellow
Pale olive-buff to yellow-ocher
_ Vinaceous-buff to light mouse-gray
None or faint yellowish
Mouse-gray, with white margin
None
Wrinkled, white Colorless to whitish, reverse cinnamon-
buff
White
| Golden yellow
Abundant, white
None
Abundant yellowish pellicle Wrinkled, yellow
White White
Golden to faint brown Faint yellowish-brown
| Rapid Medium
| White
|
Wrinkled, cream-colored Wrinkled, golden yellow to orange
White to seashell-pink
Faint brown Faint brownish
Thin, yellow pellicle
White
Golden
Colonial buff to honey-yellow
White to smoke-gray
Yellowish (golden yellow)
Rapid coagulation and peptoniza-
tion
Strong
Rapid coagulation and strong peptoni-
zation
Positive
Antibiotic production | Produces actinomycin
other morphological and cultural differences
between this culture and the two older cul-
tures, justified creation of a separate species,
especially because of the ability of S. r¢mo-
sus to produce an important new antibiotic.
In view of the great variability of these
organisms and the temptation to establish
separate species on the basis of minor differ-
ences in pigmentation, any attempt to
create such new species must be considered
critically.
Produces flaveolin
described as SS. armillatus
(Maney-Courtillet e¢ al., 1954) appeared, on
the basis of the description, to be sufficiently
close to S. rimosus to throw doubt upon its
distinct identity. Like the latter, it produced
spirals in its aerial mycelium; on synthetic
agar it formed very poor growth without any
aerial mycelium and without pigmentation;
on nutrient agar, it produced yellow-gray
growth with poorly developed white aerial
A culture
mycelium and no soluble pigment; on potato
126 THE ACTINOMYCETES, Vol. II
TABLE 13
Characterization of Streptomyces parvus (Waksman, 1957)
Characteristics | Krainsky Bergey’s 6th Edition New culture received drm N. RRL
Substrate ; Pal Golden yellow to brick-red | Bright yellow
growth |
Aerial myce- | White to gray to rose-yel-
depending on composi-
tion of medium |
| Poorly developed, rose-
Long, straight hyphae; no
spirals
| Short, oval
Thin, yellow growth; thin
lium low depending on nitro- | white; sporophores pro-
gen source duce spirals
Spores More or less oval, 1.6 w in | Spherical to oval, 0.9-1.3
| size by 1.2-1.8 pu
Synthetic | Colonies small, yellow in | Colonies small, yellow,
agar | color, with light colored) with aerial mycelium |
- aerial mycelium* light yellow |
: | : |
Nutrient Yellow growth. Aerial my-
agar celium appears late
white to yellow aerial my-
celium; bright yellow sol-
uble pigment
Yellow growth; abundant
white with grayish tinged
aerial mycelium; bright
yellow soluble pigment
Glucose as- | Yellow growth; white to
paragine | gray aerial mycelium,
agar | golden soluble pigment
Glucose agar | Aerial mycelium light yel- Colonies small, yellow, |
low; appears late with aerial mycelium |
Gelatin Colonies flat or concave,
yellow in color; gelatin |
slowly lquefied
Potato Colonies yellow, aerial my- |
celium white |
|
|
|
|
|
Cellulose | White surface growth
Remarks |
| mitrate slowly; strongly |
| proteolytic |
| Growth good
Produces diastase; reduces | Produces actinomycin; re- |
duces nitrate slightly
hight yellow
| Colonies yellow; liquefac-
tion medium
Cream-colored growth drop-
ping to bottom; good
liquefaction; bright yel-
low soluble pigment
Abundant wrinkled brown-
| ish-yellow growth; abun-
dant sulfur-yellow aerial
mycelium; no soluble pig-
ment
* Calcium malate agar
plug, it produced yellow-gray growth with a
faint brownish soluble pigment; on gelatin,
it formed a surface growth with white aerial
mycelium, with a yellowish or brownish
soluble pigment, and good liquefaction of the
gelatin; on milk, it produced good grayish
erowth. These characteristics, together with
the ability to produce oxytetracycline, defi-
nitely placed the culture in the S. riémosus
species. Emphasis was laid upon the fact it
formed flat colonies, hardly folded and not
eracked like those of S. rémosus; it showed
concentric circles in the aerial mycelium, a
variable property. It did not form nitrite
from nitrate, and it did not hydrolyze starch;
these two properties were hardly sufficient,
however, to justify the recognition of S.
armillatus as a new species.
Among the various members of the Flavus
series, the actinomycin-producers occupy an
SERIES AND SPECIES OF GENUS STREPTOMYCES
TABLE 14
Characteristics of Streptomyces flavus and allied strains (Waksman, 1957)
Characteristics S. flavus (Bergey) S. griseoflavus (Bergey) S. aureofaciens (Duggar) S. rimosus (Sobin et al.)*
Morphology
Sporophores Straight, much branched, | Straight, no spirals Straight, flexuous; no spi- | Numerous spirals
no spirals rals as a rule; occasional
loose spirals
Spores Oval Spherical to oval, 1.5 » | Short, cylindrical, 0.6-0.7
long by 0.8-1.4 4
Synthetic agar
Substrate
growth
Aerial myce-
lium
Nutrient agar
Substrate
growth
Aerial myce-
lium
Soluble pig-
ment
Gelatin
Substrate
growth
Aerial myce-
lium
Liquefaction
Soluble pig-
ment
Potato
Substrate
growth
Aerial myce-
lium
Color of plug
Yeast-glucose
agar
Substrate
growth
Aerial myce-
lium
Soluble pig-
ment
Milk
Production of
antibiotics
Yellow to sulfur-yellow
Straw-yellow
Cream-colored, lichenoid
White to light gray
None
Yellowish
None
Positive
Faint yellowish
Lichenoid, brownish to
greenish-olive
White to gray
Brownish or none
Rapid, lichenoid, brown-
ish
White, later grayish
Yellow
Coagulation and peptoni-
zation
An antibacterial
formed
agent
Reddish-brown to orange
White
Cream-colored
White
None
Cream-colored to brown-
ish-white
White
Slow
Faint yellowish
Lichenoid, brownish to
reddish-brown
White to gray
None
Cream-colored to brown-
ish
White to grayish
Yellowish
No coagulation, rapid pep-
tonization
Heavy cream-colored, be-
coming yellowish-brown.
White, turning mouse-gray
to brownish-gray
Good, light brownish
None
None
None
Wrinkled, orange-yellow
Unchanged
Heavy, cream-colored
White to deep gray or dark
gray
None
No coagulation, no pep-
tonization
Chlortetracycline
Submerged, colorless
None
Cream-colored to brownish
None or white to gray-
white
Faint yellowish or none
Moderate
White
Medium to good
Wrinkled, ochroid
Whitish to drab
Yellowish-brown
or none
pigment
| Good, yellowish
Pallid drab
Yellowish
No peptonization
Oxytetracycline and rimo-
cidin
* An earlier description of this culture on synthetic agar was incorrectly labelled; the medium was made up at that time with glu-
cose in place of sucrose, which explains the difference between the previous and present observations.
interesting place. A culture (No. 3491) be-
longing to S. flavus or to S. parvus was 1s0-
lated in our laboratories in 1948 and found
capable of producing actinomycin. It was
nonchromogenic and formed a straw-colored
to yellow aerial mycelium. This culture was
found to belong to the S. flavus subgroup;
other cultures (Nos. 3677, 3679, and 3680)
were included in the S. parvus subgroup.
Another culture (No. 3686), designated as
S. parvus, did not form any spirals. Still
another culture (No. 3687) produced only
limited curling of the aerial mycelium and
might be considered an intermediate be-
tween the two subgroups. It was suggested,
therefore (Waksman and Gregory, 1954),
THE ACTINOMYCETES, Vol. II
Fiaure 31. Variation in morphology of spore-bearing hyphae in S. aureofaciens: (left) natural vari-
ant A 377; (center) natural variant AB 374; (right) induced mutant A 377-2655 (Reproduced from:
Backus, E. J. e¢ al. Ann. N. Y. Acad. Sci. 60: 101, 1954).
that the whole series be designated as Flavus-
parvus. Considerable variation was found
among the members of this series.
Morphological Characters
Hyphae: (a) short, gnarled, and in clus-
ters, with short oval spores; or (b) long,
straight with spherical spores; or (c) long
with long corkscrew spirals and spherical
spores (Fig. 31).
Physiological Characters
Sucrose nitrate agar: Growth cream- col-
ored, yellow to brownish to orange; reverse
yellow to orange. Aerial mycelium cream-
colored, straw-colored to citron-yellow,
straw-yellow, grayish-yellow to bluish-gray
to white, or absent. Soluble pigment hght
yellow to brownish.
Glucose-asparagine agar: White to cream-
colored growth, sometimes turning orange.
Aerial mycelium white to gray. Soluble pig-
ment none, or brownish to yellow.
Nutrient agar: Growth cream-colored to
yellowish to brownish. Aerial mycelium
white, cream-colored to
Soluble pigment yellow to almost none.
Cream-colored to
eray, or absent.
Gelatin: vellow to
orange-yellow ring on surface. Aerial myce-
hum cream-colored, straw-green to gray, or
absent. Soluble pigment brownish to yellow,
or absent. Liquefaction varies from slow to
rapid.
Potato: Growth abundant, lichenoid,
cream-colored to brownish to orange. Aerial
mycelium white, cream-colored, gray to
vellow. Usually no soluble pigment; occa-
sional yellowish-orange pigment.
Milk: Surface growth abundant or thin
eray to black ring. Aerial mycelium white to
gray or absent. Milk not coagulated but
peptonized, the rapidity depending on ex-
tent of growth.
Antagonistic properties: Some members of
the group produce highly important anti-
biotics, such as the tetracyclines, that have
found extensive application in chemotherapy
and in food preservation (Kochi et al.,1952).
A careful study of the literature reveals
the fact that a large number of species found
in nature belong to this series. Some of them
have been well recognized and described.
Various others may be added, but many
have been only insufficiently described. On
the basis of the recognized information, the
S. flavus series may be said to include the
SERIES AND SPECIES OF GENUS STREPTOMYCES
following species: S. flavus, S. flavovirens, S.
flavogriseus, S. chrysomallus, S. celluloflavus,
and S. viridans.
IV. Series Ruber
Characteristic Properties
a. Sporophores — straight or — spiral-
shaped.
b. Substrate growth pink, red to red-
orange to purple-red; pigment insoluble.
Aerial mycelium thin, rose-white.
ce. Melanin-negative.
The Ruber series comprises a large, highly
heterogeneous group of organisms. Members
of this series have been known since 1888,
when Macé described an organism under
the name of Cladothrix Numerous
other cultures under different names were
later placed in this group.
The Ruber series is characterized by a
bright red, red-orange, or rose-red substrate
growth, the color depending on the composi-
tion of the medium and on conditions of
cultivation. The cultures may show con-
siderable variation in color of the substrate
mycelium, from purple-red to light rose. The
pigment is usually not excreted into the
rubra.
medium, unless the latter contains fatty sub-
stances in which the pigment is soluble. The
aerial mycelium is not well developed; it is
usually produced on synthetic media as a
thin, rose-white cover, or it is formed only
in isolated sectors or spots. The sporophores
are straight or spiral-shaped; the spores are
spherical to oval, 0.7 to 0.8 by 0.8 to 1.0 wu.
The members of the Ruber series are not
very strongly proteolytic or diastatic. Su-
crose is readily inverted. Some of the species
belonging to this series are active producers
of antibiotics.
Baldacei did not list a Ruber series, but
one designated as “‘roseus,’? which is close
enough to be considered similar to it. An-
other was designated as ‘‘melanosporeus,”’
129
which the ‘‘ruber’’ series.
Gause et al. (1957) divided the series into
is also close to
three subgroups on the basis of structure of
the namely, spiral-shaped,
straight, and tuft-forming; the last appar-
sporophores,
ently includes verticil-forming types.
Some of these organisms may be con-
sidered as forms intermediate in transition
to the true chromogenic types. Among the
forms closely related to this series, one may
inciude, for example, S. melanocyclus, S.
melanosporeus, S. melanogenes, and possibly
also S. erythrochromogenes, S. roseochromo-
The
Ruber series is also related to the Fradiae
genes, and SS. purpureochromogenes.
and Flavus series, notably through such spe-
cies as S. roseoflavus and possibly S. micro-
flavus.
Certain forms that may be considered as
species of Nocardia are frequently included
in this and in the next series. Sometimes even
a new series Is created for them, as was done
by Baldacci (1942) for ‘‘madurae.”’
S. albosporeus may be considered as a
subgroup of the Ruber series. It is charac-
terized by the formation of a rose-colored or
red to brown substrate growth and a white
aerial mycelium. Cultures belonging to this
subgroup are characterized by strong pro-
teolytic activity and by weak diastatie ac-
tion. The sporophores are straight, with
some close spirals. The first representatives
of this subgroup were isolated by Krainsky
in 1914 and by Waksman and Curtis in 1916.
The separation of members of this series
on the basis of carbon utilization has been
suggested by Zahner and Ettlinger (1957), as
shown in Table 15.
Although a large number of species found
in the literature may be included in the
Ruber series, only a few have been suffi-
ciently described. It is sufficient to mention
S. ruber, S. niveoruber, S. albosporeus, and S.
erythraeus.
THE ACTINOMYCETES, Vol. II
TABLE 15
Utilization of carbon sources by a group of closely related
Streptomyces species (Corbaz et al., 1957)
2 4 = S
Culture z z e fe » 2 z = g 3
Sluggo Ose ee 8 Fey a ie eae
“A a4 f io) 2S c 3 > T a =
De Sane Ay’ 288 oR gat ee = A nt cee
S. purpurascens a a + tr ar Seed a daiz, eee en a
S. bobiliae SI “F ls = a + she (=) C=). [Ge
S. cinereoruber + —- (-— = 2 = = Ve) ae as
S. cinereoruber var. fructofermen- + (+) + (4+) + (ey ee aS ae as
tans
+ = good growth; (+) =
— = no growth.
V. Series Viridis
Characteristic Properties
a. Sporophores straight or spiral-shaped.
b. Growth at first colorless, becoming
green to dark green. Aerial mycelium white
to gray to light green to light blue.
ce. Melanin-negative.
d. Soluble pigment absent or greenish.
The species included in this series show
considerable overlapping with the species in-
cluded in the chromogenic series, such as S.
viridochromogenes (syn. A. viridis (Lom-
bardo-Pellegrino) Baldacci).
Various other organisms that might be
included in this series have been described.
It is sufficient to list S. alboviridis, S. griseo-
viridis, and S. dassonvillet.
The following organisms may be included
in the Viridis series: S. viridis, S. prasinus,
S. hirsutus and S. prasinopilous.
Several forms described by Gause ef al.
(1957) could be ineluded in this series,
notably A. malachiticus and A. olivaceoviri-
dis.
VI. Series Violaceoruber
Characteristic Properties
a. Sporophores produce spirals. Spores
spherical to oval. Surface of spores smooth.
b. Substrate growth colorless, becoming
weak growth, questionable carbon utilization; (—) =
very weak growth;
red, later blue. Aerial mycelium white to
gray with bluish tinge.
c. Melanin-negative. Soluble red pigment
in acid media, changing to blue in alkaline.
A number of organisms belonging to the
genus Streptomyces are able to produce a
blue pigment when grown on certain media
(Tables 16, 17). This pigment is either re-
tained in the substrate mycelium or is readily
dissolved in the medium; it is frequently ac-
companied by a dark chromogenic pigment.
The color of the pigment ranges, therefore,
from light blue to dark blue or violet, and to
almost black. The soluble pigment fre-
quently changes in color with a change in
reaction of medium, from red at an acid
reaction to blue at an alkaline reaction. Be-
‘ause of this change in the color of the pig-
ment, various names, indicating the red and
blue color combinations, have been used to
describe the species, such as ‘‘vzolaceus,”
“violaceoruber,” ‘‘violaceoniger,” ‘‘tricolor,”’
and ‘“‘pluricolor.”’ The species capable of
producing blue pigments are divided here
into two distinct subgroups: S. violaceorwber
and S. vzolaceoniger. The first comprises the
forms that produce a litmus-like pigment,
changing from red in acid media to blue in
alkaline; the second includes those forms
that produce violet to dark blue to almost
violet-black pigments on synthetic and or-
b)
ganic media.
SERIES AND SPECIES OF GENUS STREPTOMYCES
TABLE 16
Organism Color of aerial mycelium
Light grayish-blue
Grayish-yellow
Greenish-grayish-yellow
Bluish-gray to blue
Greenish-brownish-gray
Gray, sometimes with
_ brownish tinge
White with purple tinge
due to substrate myce-
lium
Whitish-gray
S. caeruleus
S. coelicolor
S. cyaneofuscatus
S. cyaneus
S. cyanoflavus
S. litmocidini
S. novaecaesareae
(= A. violaceus
caesart)
. pluricolor
—R
S. tricolor
S. violaceoruber
Light brown to light gray
Ash-gray
| Ash-gray
Mouse-gray
S. olivaceus
Streptomyces sp. No.
169
—; seldom
Streptomyces species, producing a blue pigment (Kutzner and Waksman, 1959)
Melanin
pigment Author
Spirals Spore surface
= — Baldacci, 1944
_ Smooth Miller, 1908
Gause et al., 1957
Krassilnikoy, 1949
Funaki et al., 1958
Gause et al., 1957
+ Spiny
+++ 1
atts
+ = Waksman and Cur-
+ | | tis, 1916; Waks-
| man, 1919
ok Berestnew, 1897;
| Krassilnikovy,
| 1949
Wollenweber, 1920
Waksman and Cur-
tis, 1916; Waks-
man, 1919
Corbaz et al., 1957
oS Kurosawa, 1951
++
Smooth _
— Smooth =
The first organism belonging to this series
was isolated, in 1891, by Rossi-Doria and
described as Streptothrix violacea. It was later
studied by Gasperini (1894), and by San-
felice (1904) as one of the three important
constituent groups of actinomycetes.
Baldacci (1942) designated the series as
“wolaceus,” which he did not differentiate,
however, from the subgroup designated here
as violaceoniger. Gause et al. (1957) created
a new series, ‘‘roseoviolaceus,”’ which logically
belongs in this series; they also included in
their series “‘violaceus’? a variety of other
forms that logically belong to this series.
Ettlinger et al. (1958) designated as azureus
the light blue pigmented forms.
This series is not known for the production
of any important antibiotics, although coeli-
colorin has been reported for cultures of S.
violaceoruber and chartreusin for S. char-
treusts.
The following species may be included in
this series: S. violaceoruber, S. novaecaesareae,
S. cyanofuscatus, and S. litmocidini.
Some of the melanin-producing forms,
such as S. violaceochromogenes, may also be
included in this series. Many of the forms
described by Gause et al. (1957) also belong
here. These include A. coerulescens, A. glau-
cescens, A. bicolor, A.
coeruleofuscus, A. violaceorectus, A.
color, A. litmocidini, A.
griseoruber, A.
coeruleorubidus, <A.
prani-
viridoviolaceus, A.
griseorubiginosus, A. cinna-
barinus, and others.
VII. Series Fradiae
Characteristic Properties
a. Sporophores usually straight; occasional
loops and spirals.
b. Substrate growth yellow-orange to or-
ange. Aerial mycelium seashell-pink, espec-
ially on potato agar and on glucose-aspara-
gine agar.
c. Melanin-negative.
This represents a fairly large group of or-
ganisms, widely distributed in nature. S.
fradiae was first isolated and described by
TABLE
THE ACTINOMYCETES, Vol. II
17
Blue-pigmented substances produced by actinomycetes (Kutzner and Waksman, 1959)
Solubility Author
|
Preparation Organism | Melting point |
| |
|
a? a = | : i
| [c
Amylocyanin S. coelicolor | |
|
Litmocidin | Nocardia cyanea 144-146
|
|
|
‘oelicolorin S. coelicolor | 142-146
|
|
Cyanomycin | S. cyanoflavus 128
|
Granatacin S. olivaceus | 204-206
|
|
Actinorhodin S. coelicolor 270
| \
(decomp)
290-300
(decomp)
Streptocyanin Streptomyces sp. |
Anthoecyanin _S. violaceoruber
Anthocyanin S. coelicolor
Anthocyanin S. violaceoruber
Hydroactino- | Streptomyces — sp.,
chrome producing violet |
growth and pig- |
ment | |
Lipoactino- |
chrome | |
Waksman and Curtis (1916). Of the two
neomycin-producing cultures isolated by
Waksman and Lechevalier in 1949, one
formed no spirals and thus agreed with the
original description of the organism; the sec-
ond produced some spirals of the closed type.
Differences were also observed in the shade
| Very soluble in acetone, ethyl-
| Soluble in water
Inwaterandindimethylforma- | Miiller, 1908
mide; insoluble in other sol-
vents |
Shghtly soluble in water at an | Gause, 1946;
acid reaction and extracted | Brazhnikova,
from it by ethanol, ether, or | 1946
amylacetate
Kominami, 1949;
acetate, or chloroform; solu- Hatsuta, 1949
ble in ethanol, methanol,
benzene, or ether; insoluble |
in petroleum ether
Extracted from water at
alkaline reaction by chloro- |
form or methylenechloride
Extracted from water at an
acid reaction by acetone; |
soluble in ethylacetate, and |
dimethylsulfoxide; insoluble |
in petroleum ether
Soluble in pyridine, piperi-
dine, or phenol; weakly sol-
uble in dioxane or acetone;
insoluble in ether, CS: ,
CCl, or petroleum ether |
Soluble in acetone, dioxane, or | Tonolo
pyridine | 1954
Extracted with hot cold | Kriss, 1936
water and dilute alcohol
| Kriss, 1937
Funaki et al.,
1958
an |
Corbaz et al.,
1957
Brockmann et al.,
1947, 1950, 1955
Ci Males
or
Extracted with hot or cold |
water and dilute alcohol |
| Frampton and
| Taylor, 1938
Kriss, 19386
|
I
Insoluble in water
of color of the aerial mycelium on synthetic
media.
Some of the strains of S. fradiae were
found capable of producing certain antibi-
otics, notably members of the neomycin com-
plex, as well as the antifungal agent fradicin.
Several other species reported in the litera-
SERIES AND SPECIES OF
ture appear to be related to the Fradiae
series. Baldacci et al. (1953) at first did not
recognize this organism as representing a
distinct series, and apparently considered it
as a within the
Later, Baldacci
(1956) gave it series characteristics.
Gause et al. (1957) divided the Fradiae
member “roseus”’ series.
however, and Comaschi
series, on the basis of spiral formation, into
two subgroups: one, spiral-forming, com-
prising A. roseoflavus; the other, nonspiral-
forming, comprising A. fradiae proper; other
species and varieties were included in both
subgroups. Most of the members of a new
series, designated as ‘‘fuscus,’’? could also be
considered as members of the /’radzae series.
Waksman and Scotti (1958) divided the
Fradiae series into three subgroups. These
were described briefly as follows:
I. Substrate growth on synthetic media
thin, smooth, colorless, almost entirely lim-
ited to the surface of the medium; occasion-
ally colored orange-yellow. Aerial mycelium
light pink, seashell-pink, or salmon-colored.
Some strains produced little if any growth
on synthetic media. Best sporulation took
place on potato agar and on glucose-aspara-
gine agar. On organic media, growth was
smooth to wrinkled, yellowish or orange-
vellow to orange-brown,; aerial mycelium, if
present, was white to seashell-pink. On cer-
tain media, a soluble, pink to salmon-colored
pigment was produced. Morphologically, all
strains formed a straight aerial mycelium;
some cultures, however, were able to form
hooks and loops, and even occasional spirals,
on certain media. These strains were con-
sidered as representing typical S. fradzae
proper.
Ila. On synthetic media, substrate growth
thin, colorless, limited almost entirely to the
surface of the medium; aerial mycelium
white. On organic media, growth cream-
colored to yellowish; aerial mycelium thin,
white to grayish-white. On yeast-glucose
agar, growth orange to brownish to greenish;
GENUS STREPTOMYCSE 133
aerial mycelium white. Abundant spirals
were found in the aerial mycelium.
IIb. On synthetic media, growth very
poor. On organic media, growth generally
poor; growth best on yeast-glucose agar.
No aerial mycelium was formed.
A detailed characterization of subgroup I
is given in Table 18. Among the other species
apparently closely related to this section is
S. kanamyceticus (Okami and Ume-
ZAWAa).
The following species may be included in
the Fradiae series: S. fradiae, S. luridus, S.
albosporeus, perhaps also S. roseus and S.
fuscus.
VILL. Series Griseus
Characteristic Properties
a. Sporophores straight, produced in tufts.
Spores oval; surface smooth.
b. Growth colorless to olive-buff. Aerial
mycelium water-green to grass-green to gray.
c. Melanin-negative.
d. Strong proteolytic activities. Produce a
variety of antibiotics.
An organism, under the name of A. griseus,
was first isolated and described by Krainsky
in 1914. Its substrate growth on artificial
media was colorless; only a small amount of
yellowish soluble pigment was produced. The
aerial mycelium was of a green-gray color
on both organic and synthetic media. When
the concentration of nitrogen in the medium
was increased to 0.005 per cent, the aerial
mycehum became white. The culture was
only weakly proteolytic.
Soon afterward, in 1915, Waksman and
Curtis isolated several cultures of what ap-
peared to be the same organism, the com-
parison with Krainsky’s description being
based primarily on the color of the aerial
mycelium. Since this work was done during
the of World War I, Krainsky’s
original strain could not be obtained for com-
years
parative studies. The new culture was des-
THE ACTINOMYCETES, Vol. II
154
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SERIES AND SPECIES OF
ignated as A. griseus Krainsky, although
certain marked differences were observed
between the two isolates.
Since no type culture of Krainsky’s organ-
ism was available for comparison to any
investigator, all the subsequent descriptions
were based upon the Waksman and Curtis
culture, which was distributed to all collec-
tions in the world.
In 1919, Waksman amended the descrip-
tion of Krainsky, as follows: ‘‘This organism
was isolated numerous times from the soil.
The name A. griseus was used before by
Krainsky so that the description of the latter
is itself an amendment. Although this or-
ganism was originally identified with the
organism described by Krainsky under the
same name (from description only, without
any actual comparison of cultures), this
identification should be, therefore, corrected.
The culture described here possesses a very
strong proteolytic power, while Krainsky
stated that his culture was not strong pro-
teolytically.”’
The differences between the two cultures
can be briefly summarized as follows: A.
griseus Krainsky produced a greenish-gray
to dark gray aerial mycelium, with a green-
ish-yellow soluble pigment in older cultures;
growth on potato was grayish, with white-
gray aerial mycelium; Krainsky never stud-
ied the morphology of his organism, except
for the shape (oval) and size of the spores.
A. griseus Waksman and Curtis produced a
water-green to yellowish-green aerial myce-
lium; the sporophores were straight and were
formed in tuft-like masses; growth on potato
was yellowish, wrinkled, and without any
soluble pigment.
Tn spite of these differences, Waksman
hesitated at first to change the name of the
culture which he and Curtis first isolated.
This hesitation was due partly to the faci
that the organism was found to undergo
considerable variation upon continued cul-
tivation on artificial media. The substrate.
GENUS STREPTOMYCES
the temperature of incubation, the length of
the incubation period, the amount and na-
ture of inoculum, all tended to exert an in-
fluence upon the morphological and cultural
characteristics of the organism. At one time
milk was clotted at 37°C in 2 days and then
peptonized; at another time, under the same
conditions, clotting of the milk required 5
to 6 days; at still another time, the milk
in some tubes was not clotted at all but was
rapidly peptonized. There were other recog-
nizable changes or Drechsler,
studying the morphology of the Waksman
and Curtis culture, found that the aerial
mycelium showed proliferation of fertile
branches at moderately close intervals along
the axial hyphae, thus suggesting tuft for-
mation. This phenomenon alone would have
definitely indicated that the culture should
have been identified as a distinct species.
In August 1943, in the laboratories of the
Department of Microbiology of the New
Jersey Agricultural Experiment Station, a
culture was isolated which produced the
highly important antibiotic designated as
streptomycin. Upon careful examination,
this culture was found to be similar to the
Actinomyces griseus described by Waksman
and Curtis in 1916. Since, in the meantime,
Waksman and Henrici had proposed that the
generic name for the sporulating forms of
variations.
actinomycetes be changed from Actinomyces
to Streptomyces, the organism was named
Streptomyces griseus. This name has been
universally recognized, since 1944, as the
official one for the streptomycin-producing
organism, and has been so designated in nu-
merous throughout the
world. A detailed description of this species
was published in 1948 (Waksman, Reilly
and Harris).
Baldacci et al. (1954) subjected the Griseus
series to a detailed study. They recognized
other collections
that this representative species had come to
the fore as a result of the important role
that it played in the production of antibi-
156
otics. They emphasized that although first
listed by Krainsky in 1914, S. griseus was
amended and described in detail by Waks-
man in 1919. They further added:
“Tf we examine the characteristics given
by Krainsky we are led to link this species
with A. viridis. This conclusion appears still
more logical when we study the coloured
tables prepared by Krainsky. However, in
view of the impossibility of comparing
Kkrainsky’s original strain and the difficulties
that would arise if one did not accept Waks-
man’s amendment for a species so generally
studied in laboratories, 1t appears advisable
to take as definite the characteristics speci-
fied by the American author and given in
Bergey’s Manual .... Numerous strains have
been isolated by us and compared, with
satisfactory results, with Waksman’s strains.
_..There is a considerable body of literature
dealing with this species which has a faculty
for mutation.”
Baldacci and Comaschi later (1956) wrote:
“The examination of Krainsky’s descrip-
tion and colored pictures would suggest that
this species belongs to A. virzdis Lombardo
Pellegrino (1903). The comparison between
Krainsky’s and Waksman’s descriptions
gives evidence—as even Waksman has par-
tially pointed out—to the difference of
proteolytic activity and. according to our
opinion, the very important difference of the
color of the sporulating colonies which are
greenish in Krainsky’s description. If we ac-
cept Waksman’s correction of the species
and compare his descriptions with our
strains, we find a perfect identity. Since it is
impossible to compare the original strain of
Krainsky with the others, the acceptance of
the correction proposed by Waksman offers
the advantage of maintaining the name
“griseus” for an actinomycete so largely
spread out and studied in laboratories, so
that we agree with it according to this mean-
ing.”
Many other cultures of S. griseus have
THE ACTINOMYCETES, Vol. II
since been isolated from soils, river muds,
animal excreta, water, dust, and other nat-
ural substrates. Not all of them were found
capable, however, of producing streptomy-
cin; the majority of these cultures were
either inactive or produced other antibiotics,
such as cycloheximide, grisein, streptocin,
actinomycin, and candicidin. Some of the
cultures yielded a mixture of streptomycin
with other antibiotics. The ability to form
streptomycin was at first considered as a
strain, rather than a species, characteristic;
later, however, it was decided (Waksman,
1959) to raise S. griseus to the status of a
the — streptomycin-producing
strains to a species status, Streptomyces
griseus, Waksman and Henrici.
Several procedures were developed for the
isolation from natural substrates and for the
identification — of
strains of S. griseus. These methods were
based on certain physiological properties of
the organisms and on the nature and ac-
series and
streptomycin-producing
tivities of the streptomycin formed by them:
1. Tolerance to fairly high concentrations
of streptomycin in the medium. When a
soil or other natural material was plated
out on a medium containing 50 mg of strep-
tomycin per liter, the great majority of bac-
teria and actinomycetes failed to develop on
the plate. Most of the actinomycete colonies
were found to be of the S. griseus type.
2. Ability of certain resistant strains of
test bacteria to grow in the presence of
streptomycin.
3. Sensitivity to a specific actinophage.
When cultures of S. griseus are tested for
their sensitivity to a specific actinophage
which is active only upon the streptomycin-
producing species, the inactive forms or those
producing other antibiotics can be easily
eliminated.
4. Utilization of streptomycin-dependent
strains of bacteria in testing for strep-
tomycin. When a culture of S. griseus or of
another organism suspected of producing
SERIES AND SPECIES OF GENUS STREPTOMYCES 137
streptomycin was finally selected and grown
in a liquid medium, the streptomycin-like
nature of the antibiotic could be established
by adding the culture filtrate to a nutrient
broth and inoculating the latter with a strep-
tomycin-dependent strain of Escherichia coli
or of some other bacterium. Growth of the
bacterium definitely established the fact that
the unknown antibiotic was streptomycin.
5. Cross-streaking the unknown cultures
on a suitable agar medium toward known
streptomycin-producing cultures. The latter
exerted only a shght inhibiting effect upon
the unknown streptomycin-producers.
Usually some soil or other material is
plated on ordinary agar media favorable to
the development of actinomycetes; colonies
were picked and tested. The S. griseus col-
onies could easily be recognized by the pale
green to grayish-green shade of their aerial
mycelium. A suitable agar medium can also
be seeded with living cells of a nonpatho-
genic strain of Mycobacterium tuberculosis
and various dilutions of soil used for plating
purposes. The plates are first incubated at
28-30°C for 2 or 3 days, to enable the acti-
nomycetes to develop. This is followed by
further incubation of the plates at 37°C for
the development of the test bacterium. Col-
onies that have the capacity of inhibiting
growth of the bacterium are found to be
surrounded by clear zones.
The antibiotic potency of an active culture
of SS. griseus was found to be fairly constant,
in spite of the ability of the culture to give
inactive variants. Highly active
strains tend to retain their relatively superior
streptomycin-producing potency,
poor strains usually remain weak producers
of this antibiotic. For the commercial pro-
duction of streptomycin, however, it is es-
sential to select continuously the most ac-
rise to
whereas
tive strains.
Since the streptomycin-producing culture
isolated in 1943 was found to be identical
with the one described by Waksman and
Curtis, 1t must be considered in the light of
that description. The same is true of the S.
griseus strains isolated later and found to
be able to produce grisein, candicidin, vio-
mycin, and actinomycin. There are certain
differences in the cultural and biochemical
properties of the various strains belonging
to the Griseus series, especially in their ability
to produce various antibiotics and in their
sensitivity to different phages. This justifies
the separation of the group into several dis-
tinct species.
The morphological and cultural properties
of certain cultures belonging to the Griseus
series are given in Table 19.
Characterization
The Griseus series is characterized by cer-
tain morphological and cultural properties
that make possible its identification and
ready distinction from other groups belong-
ing to the genus Streptomyces. As more and
more cultures of S. griseus were isolated, it
became recognized that this is a large series
of organisms, the members of which vary
greatly in their physiological properties and
in their ability to produce various antibiot-
ics.
Waksman and Curtis described S. griseus
as producing on sucrose nitrate agar a thin,
spreading growth, developing deep into the
medium, at first colorless, then turning olive-
buff. This color may be lost on successive
transfers. The aerial mycelium is thick, pow-
dery, water-green in color. No soluble pig-
ment was observed; the reverse of the growth
became brownish in 24 days. On gelatin,
at 18°C, S. griseus produced a greenish-yel-
low or cream-colored growth developing deep
into the substrate; the aerial mycelium was
white-gray with a greenish tinge. There was
no soluble pigment; liquefaction of the gela-
tin was rapid. The culture was capable of
utilizing a variety of carbohydrates, includ-
ing pentoses, hexoses, sugar alcohols, and
organic acids. It was also able to obtain its
138
THE ACTINOMYCETES, Vol. II
TABLE 19
Morphological and physiological properties of certain strains and one mutant of Streptomyces griseus
| Synthetic agar | Glycerol agar Gelatin* Potato | Anebi
ea): | | 2
Se An eENO aL yeep ar TG | Sea eee:
Growth oe celta | Growth eyes Growth aves ties
—— —|—— = i— - | |
1915 W and C | Sporophores Cream-col- | Powdery, | Cream-col- White to | Cream-col- White to | Of
(N. J. strain) long, formed ored water- | ored | greenish- ored or greenish- |
in tufts, no | green yellow greenish- yellow
spirals color yellow
1915 W and C Sporophores Cream-col- Powdery, | Cream-col- White to | Greenish- Grayish ++
(Holland long, formed ored, turn- water- ored greenish- | yellowor |
strain) in tufts, no ing olive- green yellow cream - |
spirals buff color | | colored |
1943 Strepto- Tufts, no Cream-col- Powdery, Cream to Cream-col- | Greenish | Greenish |-++-+-+
mycin pro- spirals ored, turning water- olive- ored yellow or | tinge
ducer olive-buff green green cream-
color colored |
Grisein pro- Tufts, no Cream_-col- | Light gray Cream-col- Cream-col- | Greenish- | Grayish } ++
ducer spirals ored to green- ored ored to yellow
ish | greenish |
Rhodomycetin | Tufts, no | Vinaceous White to | Carmine- Gray +++ | Gray +
producer spirals gray red | | |
| |
* Brownish pigment produced by some strains.
+ Mutant of this culture produced an antibiotic.
nitrogen from a variety of compounds, in-
cluding both inorganic and organic forms.
In studies of streptomycin-producing
strains, Carvajal (1946) characterized the
morphology and life cycle of S. griseus in
greater detail. The substrate mycelium when
young is well branched, typically in a mon-
opodial form. Transverse septa are formed
in virtually all cases in the delimitation of
the reproductive cells. Reproduction occurs
by means of unicellular asexual’spores and
conidia, which are exogenously borne in
chains on the aerial mycelium. The spores
are of various shapes: barrel, oval, bean,
spherical, and cylindrical. Differences in
shape and size are found often, even
among the spores of the same chain. Mature
aerial spores often show small fragments of
transparent film adhering to the outside wall.
The spores germinate at one end or at both
ends, usually from the points at which they
are attached to the adjacent spores or to the
hypha. Hyphal fusions and germ tube fusions
also can be observed. Carvajal reported that
he had demonstrated a nucleus in the germ
tubes of S. griseus in the young mycelium
and in the developing spores. The nuclei
were said to be well distributed throughout
the cytoplasm of the mycelium; the spores
may be uninucleate or multinucleate.
Gottlieb and Anderson (1947) studied the
course of spore germination and of develop-
ment of the mycelium in submerged cultures
of S. griseus. The exact time of spore germi-
nation was difficult to determine, only an
elongation of the spores being observed.
After 6 hours, the mycelium was found to
consist of some small hyphae and of longer
branched hyphae which tended to develop
into masses of mycelium consisting of a
dense solid center and a_ periphery of
branched radiating hyphae. Within 24 to
30 hours, the entire body of the medium
was filled with these mycelial clumps. The
culture appeared viscous at this stage. After
48 hours, the mycelium began to fragment,
and spores were produced. At 84 hours, det-
inite lysis of the mycelium took place; the
dense central core of the masses of growth
disintegrated into granular pieces.
Measurement of viscosity and weight of
mycelium revealed an increase which reached
SERIES AND SPECIES OF
a maximum at 24 to 30 hours, followed by
a decrease up to about 96 hours; a gradual
leveling of growth then took place.
Growth of S. griseus in stationary cultures
reaches a maximum in 10 days, whereas
maximum growth in submerged cultures is
usually attained in 3 to 5 days. This is fol-
lowed by lysis of the mycelium. Growth of
the organism is accompanied by a gradual
rise in pH value of the culture and in the
ammonia and amino nitrogen content; the
total nitrogen in the mycelium tends to be
higher during the active stages of growth.
The production and accumulation of strep-
tomycin parallel the growth of the organism,
reaching a maximum when lysis just sets in;
this is followed by a decrease when the rate
of lysis reaches a maximum.
Metabolism
The metabolic changes of S. griseus in a
elucose-peptone-meat extract medium have
been found by Dulaney and Perlman (1947)
to fall into two phases. During the first
phase, the organism grows rapidly and forms
extensive mycelium; this is accompanied by
a reduction in the quantity of soluble con-
stituents in the medium, namely, the nitro-
gen, the inorganic phosphate, and the avail-
able carbohydrate; the quantity of lactic
acid present is first increased and then uti-
lized to some extent; the oxygen demand is
high, and the Qo. values may reach 150;
little streptomycin is produced; the soluble
‘carbon content of the medium during the
growth phase rapidly falls as the glucose is
utilized; about 50 per cent of the carbon
appears to be unavailable to the organism
during the first stage; the nitrogen content
of the mycelium varies with age. During the
second or autolytic phase of growth con-
siderable lysis sets in; streptomycin is pro-
duced actively, and the pH of the medium
rises; the quantity of mycelium is decreased
as a result of lysis; the lactic acid content
remains more or less constant, as does the
GENUS STREPTOMYCES 139
soluble carbon content of the medium; the
oxygen demand slowly decreases; the am-
monia nitrogen, soluble nitrogen, and inor-
ganic phosphate contents of the medium
rise rather markedly, paralleling the autoly-
sis of the cells.
Ammonium compounds, but not nitrates
are favorable sources of nitrogen for growth
and streptomycin production. S. griseus
rapidly assimilates phosphate in a phos-
phorus-poor medium. An excess of phos-
phorus has a depressive effect both upon
growth of the organism and upon strepto-
mycin production.
The supplementary addition of amino ac-
ids or of more complex organic compounds
has been found to stimulate production of
streptomycin. Kiser and McFarlane (1948)
found that, of the amino acids, histidine is
essential for both mycelial growth and strep-
tomycin production; inositol also increased
the yield of both; valine favored the latter,
and aspartic or glutamic acid the former.
If the salt concentration is low, most of the
streptomycin will be found in the mycelium,
thus suggesting that streptomycin is a prod-
uct of intracellular synthesis. Woodruff and
Ruger (1948) reported that yields as high as
1 g of streptomycin per liter are produced
by S. griseus in media containing proline
as the only source of nitrogen.
The ability of S. griseus to form an en-
zyme (mannosidostreptomycinase) which de-
composes mannosidostreptomycin into strep-
tomycin and mannose has been recently
demonstrated. This enzyme is not produced
by other actinomycetes or fungi (Volume I,
p18):
On a dry basis, the mycelium of S. griseus
contains about 16 per cent ether-soluble
material and about 37 per cent cold water-
soluble substances. Little study has been
made of the specific chemical composition
of these and other fractions.
Stokes and Gunness (1946) grew S. griseus
in stationary cultures in a nutrient medium
140
containing 0.5 per cent meat extract and 1
per cent glucose. The cell material was dried
and then hydrolyzed by acid or alkali. The
amino acid composition of this material, on
a percentage basis of the dry material, was
as follows: total nitrogen, 9.14; histidine,
0.84; arginine, 2.90; lysine, 2.13; leucine,
3.73; isoleucine, 1.49; valine, 3.40; methio-
2.33; phenylalanine,
nine, 0.55; threonine, 2.33;
1.67; tryptophan, 0.62.
In addition to the two forms of strepto-
mycin, S. griseus produces several other
antibiotics. Ether extracts from the myce-
lium of the organism yield a substance
designated as streptocin, which is active
against bacteria but not
against gram-negative forms. Another anti-
biotic, designated as cycloheximide, can be
isolated by extracting the crude submerged
eulture with chloroform, evaporating the
extract, and dissolving the residue in meth-
anol. Cycloheximide is not active against
bacteria but has strong antifungal prop-
erties; it is particularly active against yeasts,
gram-positive
Varieties and Mutants
The Griseus series represents a large?
widely distributed, and variable
organisms. It has long been recognized
(Waksman, 1959) that this series should be
divided into several species. The formation
of different antibiotics by the various species
offers an excellent supplementary basis for
such subdivision. The many cultures isolated
and studied in detail can thus be classified
into five distinct species.
1. Streptomyces
group. ot
Waksman and
Henrici. This comprises strains of S. griseus
which produce streptomycin; they also pro-
duce cycloheximide.
2. Streptomyces
QriSeUs
Waksman.
Strains of S. griseus which produce grisein or
griseinus
erisein-like substances. These strains are as a
rule resistant to actinophage. Benedict and
Lindenfelser (1951) demonstrated that a
majority of streptomycin-producing strains
THE ACTINOMYCETES, Vol. II
of S. griseus form a green soluble pigment
in calcium malate medium and a_ yellow
pigment in calcium succinate medium; on
the other hand, the grisein-producing strains
of this organism do not form any green or
yellow pigments in these media, although
they show the typical greenish pigmentation
of the aerial mycelium.
3. Streptomyces coelicolor (Miller) emend.
Kutzner and Waksman. This species com-
prises strains which produce the antifungal
agent candicidin but no antibacterial sub-
stance. The first organsim belonging to this
species was Isolated by Miiller (1908), and
designated Streptothrix coelicolor. It produced
a blue pigment similar to that formed by a
diphtheroid organism which he called Bacil-
lus coelicolor. The culture was a_ typical
Streptomyces and formed concentric rings in
its aerial mycelium. It developed well at
room temperature and at 36°C. It grew on
gelatin, with gradual liquefaction but with-
out pigmentation. On agar media containing
5 to 10 per cent dextrin, but not in glycerol
media, a brown pigment was formed. The
culture formed no aerial mycelium on ordi-
nary agar media, unless serum, glycogen,
dextrin, or starch was added. When glucose,
sucrose, arabinose, or other sugars were used,
no aerial mycelium was formed.
Miller emphasized two important activ-
ities of S. coelicolor: It possessed antagonistic
properties, and it was active against Ozdium
lactis. Miller was thus one of the first to
demonstrate activities that were to make the
whole group of actinomycetes famous. Miil-
ler also studied the pigment extensively; he
called it amylocyanin; it is produced best on
potato media when grown at 30°C, but not
at 36°C.
4. Streptomyces californicus Waksman and
Curtis. This species comprises strains which
produce viomycin, active against gram-posi-
tive bacteria only. Burkholder et al. (1955)
classified these organisms as strains of S.
Waksman (1958)
griseus var. purpureus.
TT
SERIES AND SPECIES OF GENUS STREPTOMYCES
TABLE 20
141
Comparison of cultural characteristics of four strains of Streptomyces californicus with Streptomyces griseus on five different media
(Burkholder et al, 1955)
Agar medium | Mycelium S. floridae S. californicus S. puniceus S. vinaceus S. griseus 3475
Glycerol as- | Substrate | White to light White to light Purple White to light White to light yel-
paragine | gray to slight | gray to purple gray to slight low
purple purple
Aerial White White White to light White to light White to light
gray-green gray-green gray-green
Glucose tryp- | Substrate | Black with slight| Gray to slight | Gray to black Gray to black Light gray-yellow
tone purple tinge black
Aerial White to slight Slight growth, | Light gray-pink Light gray-pink White to gray-pink
| gray-pink | white to light gray- | toslight light
green | gray-green
Starch syn- Substrate | Purple Light gray to slight! Purple White tolightgray | Light yellow-gray
thetic | purple to slight purple
| Aerial White to light | White White to light | Light gray-green | Light gray-green
gray-green gray-green
Calcium malate) Substrate | White to light White to light | White | White | Gray-yellow to
yellow yellow | slight light
| | brown
| Aerial White White | Light gray-green | White White to slight
| gray-green
Nutrient | Substrate | White to slight White to slight | White to light | White to light Light gray-yellow
light yellow light yellow yellow | gray-yellow
Aerial White | White White White to slight White to slight
| light gray-green| gray-green
suggested that they be raised to the status
of a species within the S. griseus section. A
further study of this species points to its
identity to S. californicus, which has _pri-
ority in species designation.
5. Streptomyces chrysomallus (Lindenbein)
Waksman. These comprise strains which
produce actinomycin. Welsch et al. (1957)
studied 51 cultures of Streptomyces for their
susceptibility to seven actinophages; 43
of the strains produced actinomycin and
eight represented nonactinomycin-producing
strains of S. griseus. Certain actinomycin-
producing organisms, including the Linden-
bein culture of S. chrysomallus and a culture
of S. parvus, were considered to belong to
the S. griseus section. A detailed study of the
actinomycin-producing organisms has _ re-
cently been made by Solovieva and Die-
lova (1960).
Various other organisms belonging to the
S. griseus section are able to form at least two
other antibiotics. One of these, cyclohex-
imide, is active only against fungi, and
another, streptocin, is active against certain
protozoan-like organisms. No detailed study
has as yet been made of these strains in an
effort to raise them to species status.
There are also those strains of S. griseus
that produce no antibiotic at all, at least as
far as one is able to detect by available
methods.
The streptomycin-producing strains of S.
griseus give rise readily to mutants. Two
such mutants have been reported: One was
a colorless form, producing no aerial myce-
lium, forming no streptomycin, and sensitive
to this antibiotic; Dulaney et al. (1949) re-
ported, however, on a colorless mutant that
produced streptomycin. The other was a
pigmented mutant, forming pink to vina-
ceous-colored substrate growth and an aerial
mycelium typical of S. griseus; this mutant
formed no streptomycin but gave rise to
another
was not
teria. According to Kutzner (1960), this
antibiotic (rhodomycetin), which
active against gram-negative bac-
strain shows much similarity, on the basis of
phage sensitivity and other properties, to
S. californicus. A detailed study of degenera-
142 THE ACTINON
IYCETES, Vol. II
TABLE 21
Classification of actinomycin-producing organisms based upon utilization of rhamnose and
raffinose (Bt
tlinger et al., 1956)
Utilization of:
Actinomycin Species Reference or origin of strain
Rhamnose Raffinose
+ S. flavovirens Pridham and Gottlieb + +
+ S. flaveolus Kurosawa + +
oo S. antibioticus Pridham and Gottlieb + =
+ S. antibioticus Burkholder e¢ al. + =
xX S. antibioticus orig. (Waksman) + =
XE S. antibioticus NRRL (Raper) + —
C S. chrysomallus orig. (Lindenbein) + =
_ S. flaveolus orig. (Waksman) + =
I S. parvullus orig. (Waksman) + —
xX S. parvus NRRL (Benedict) + —
x 3 ETH strains _
C 8 ETH strains a -
x YTH 9001 _ _
TABLE 22
Grouping of actinomycin-producing organisms (Ettlinger et al., 1956)
Group arena Spore color ee Authentic strains ETH strains
I + cinereus x S. antibioticus (orig. and 33
NRRL)
II + griseus 1
. S. parv s (orig. al
III is mcans ' parvullus (orig.) :
( S. chrysomallus (orig. ) 17
IV — griseus is)
[ S. parvus (NRRL)
tion and regeneration of S. griseus was made
by Williams and McCoy (1953).
Different varieties of S. griseus vary
greatly in their cross-resistance and in their
sensitivity to actinophages.
It must further be noted that high-yielding
streptomycin strains can be obtained by
irradiation, by growth in media containing
increasing concentrations of streptomycin,
and by strain selection.
Okami (1950a) examined 47 strains be-
longing to the Griseus series. Five of these
were grisein-forming strains and four were
pink pigmented forms. The streptomycin-
producing strains grew in maltose-containing
media with NaNQOs: as a source of nitrogen,
but not in glucose, glycerol, or sucrose media.
The grisein strains grew in media containing
any of the four carbon compounds. The pink
strains grew only in glycerol media (Table
23). The utilization of the carbohydrate was
found to depend largely on the nitrogen
source. In the presence of ammonium sulfate,
the above differences disappeared. All strains
utilized xylose, but not raffinose or rham-
hose. Sensitivity to phage was said to be
SERIES AND SPECIES OF GENUS STREPTOMYCES
strain specific, but not characteristic of
streptomycin production. The use of strep-
tomycin-resistant and — streptomycin-de-
pendent cultures of bacteria as test organ-
isms for the differentiation of the various
strains of S. griseus was considered as supple-
mentary to the foregoing differentiation
methods.
It is of particular interest, in this con-
nection, to draw attention to the confusion
that has arisen in some cases from Krassil-
nikov’s attempt to change the name of the
streptomycin-producing organism. Just as
Waksman did previously, Krassilnikov came
to the conclusion that there is a difference
between the Krainsky and the Waksman
and Curtis cultures of A. gréseus. Although
he, likewise, had no opportunity to compare
Krainsky’s original isolate with the strep-
tomycin-producing organism, he attempted
to draw conclusions on the basis of cultures
that he isolated himself, and proposed that
the name of the streptomycin-producing
organism be changed to Actinomyces globi-
sporus streptomycint (later changed to A.
streptomycina). This suggested change was
most unfortunate for several reasons: (a) a
well described specific name, namely Strep-
tomyces griseus Waksman and Henrici, was
set aside merely for the sake of priority of a
name of a culture (A. griseus Krainsky)
which no one had ever seen and which was
not available in any culture collection; (b) a
hame of an organism that had become rec-
ognized throughout the world because of its
important physiological and biochemical
properties, and especially because of its
capacity to produce a highly important
chemical substance, streptomycin,
changed to a trinomial merely because of the
existence of an insufficiently described vari-
ety of an unknown culture.
The confusion thus became compounded
by this attempt to change the name of the
streptomycin-producing organism. We find,
in addition to the two names suggested by
Was
145
TABLE 23
Utilization of carbon sources by various strains of
Streptomyces griseus with nitrate as source of
nitrogen (Okami, 1950)
Garbonsource Streptomycin Grisein Pink-pig-
strain strain ment strain
Glucose - + —
Glycerol — + +
Sucrose _ te —
Maltose + + =
KXrassiinikov, the incorrect names Actino-
myces griseus Waksman listed by Koreniako
and Nikitina (Shorin, 1957), Streptomyces
griseus Krassilnikov by Znamenskaia et al.
(1957), Actinomyces griseomycini by Gause
et al. (1957), and finally Streptomyces glo-
bisporus streptomycini by Severin and Gor-
skaia (1957). This confusion was fortunately
limited to the literature published in Rus-
sian.
For the reasons presented here, Krassil-
nikov’s modifications of the name S. griseus,
with all the subsequent confusing names,
cannot be accepted. In fact, the actual
culture, A. (Krainsky )
Krassilnikov (1949), belongs rather to the
Cinereus series.
Qriseus emend.
The logical name for the streptomycin-
producing
griseus Waksman and Henrici.
species remains Streptomyces
Additional Organisms
Numerous other organisms belonging to
the Griseus series have been described as
species and as varieties. Some of the descrip-
tions are incomplete; and it is, therefore,
rather difficult to give them an exact posi-
tion. This is true, for example, of the cultures
described by Gause et al. (1957) under the
name A.
other species placed in the series /Helvolus.
See also Harada, 1959.
rubiginosohelvolus and some of the
IX. Series Hygroscopicus
This series comprises organisms that form
a white to gray aerial mycelium, with a
144
tendency to become dark gray; frequently
black patches are produced in the mycelium,
the whole often becoming black. The sub-
strate growth is dark gray with a tendency to
become moist, slimy, and finally changing
to black. The species are melanin-negative,
although on synthetic agar a brown to black
soluble pigment may be produced.
Morphologically the species give rise to
spiral-shaped sporophores. This series com-
prises a number of species, some of which
are listed: S. endus, S. hygroscopicus, S.
limosus, S. nigrificans, S. platensis, and S.
violaceoniger.
X. Series Scabies
Characteristic Properties
a. Sporophores produce spirals.
b. Aerial mycelium lght gray to dark
eray.
c. Melanin-positive.
d. Some species are able to cause diseases
of plants, notably scab of potatoes.
The melanin-producing capacity of certain
actinomycetes, or their ability to form solu-
ble brown to black pigments when grown in
protein-containing media, was first recog-
nized as a diagnostic characteristic by Rossi-
Doria and Gasperini in 1891. Numerous
cultures found capable of producing such
pigments were isolated from different sub-
strates, and designated as Streptothrix chro-
mogena, Actinomyces chromogenus, or <A.
chromogenes. Gradually it came to be recog-
nized that all these isolates represented not
a single species but a large number of organ-
isms, differing greatly in their morphological,
physiological, and biochemical properties.
This was definitely established in 1900 by
Beijerinck, who isolated two types of actino-
mycetes (‘‘Streptothrix”’) represented abun-
dantly in nature. “‘One of these that I have
learned to recognize in the form of numerous
varieties, I will designate as Str. chromogena
Gasperini, since I believe that one such
THE ACTINOMYCETES, Vol. II
variety was available to the author of this
name. The other species I designate as Str.
alba.” The first was characterized by the
formation of a brown pigment in meat ex-
tract-gelatin media.
Neukirch (1902) demonstrated the pres-
ence in nature of two chromogenic types of
actinomycetes. Krainsky (1914) described
four chromogenic species, Whereas Waksman
and Curtis (1916) demonstrated the occur-
rence in soil of various other chromogenic
types, “each with such well defined charac-
ters as to make it almost impossible to
classify them as one species.”
One of the most important series among
the chromogenic actinomycetes is S. scabies,
which at one time was designated as S.
chromogenus. In addition to the members of
the Chromogenus series as such, numerous
other species now included in other groups
also possess chromogenic properties, espe-
cially members of the Lavendulae and Re-
ticult series.
The soluble pigment produced by various
organisms when grown on protein-containing
media was found to belong to the melanin
type. It frequently involved the tyrosinase
reaction. The intensity of the pigment varies
with the organism and with the medium.
The formation of the melanin pigment is
usually determined by growing the organ-
isms on tyrosine-containing media.
Baldacci et al. (1953) did not recognize a
“Chromogenus” series as such, although they
Gause et al.
listed one under ‘‘Cas-Gri.”
(1957) listed two series, one a “Chromogenus”’
proper, and the other ‘‘Helvolus,” in which
both pigment-producing and nonpigment-
producing types are included.
The organisms belonging to S. scabies were
at first believed to be primarily associated
with scabbiness in white potatoes, sugar
beets, and mangels. Only the typical chro-
mogens were at first included in this series.
They formed a brown to dark brown to black
growth, a gray aerial mycelium, and a brown
SERIES AND SPECIES OF GENUS STREPTOMYCES
to black soluble pigment on organic media.
It must be conceded at once that not all
organisms isolated from scabby potatoes or
beets are able to produce a soluble brown
pigment and certainly not all are capable of
causing scabbiness in potatoes.
Millard and Burr (1926) isolated a num-
ber of cultures from scabby potatoes and
beets. They proposed a key for the identifica-
tion of the presumably potato-disease-pro-
ducing actinomycetes (see Volume I, Chap-
ter 18). The medium this
purpose (glycerol nitrate solution) could
hardly be considered the most desirable sub-
strate for bringing out the proper characters
for a system of classification. Some of these
cultures, notably S. clavifer and S. fimbri-
atus, definitely belong to the Scabies series.
Baldacci and Spalla (1956) suggested that
the strain of S. scabies isolated by Millard
and Burr be designated as S. scabies var.
anglica. It is distinguished from the North
American species first described by Thaxter
as having a “gray substrate growth, a gray
aerial mycelium and a yellow soluble pig-
ment.”
The possibility that different strains or
races of SS. scabies were responsible for the
infection of potatoes and mangels has been
fully recognized. No definite correlation has
been found, however, between pathogenicity
and cultural and other properties of the or-
ganism, although variants may differ from
the parent culture in pathogenicity. High
nitrogen content of the medium appeared to
inhibit production of aerial mycelium in the
parasitic strains but not in the saprophytes.
Of the 20 isolates tested by Schaal (1944) on
three different media, six did not produce
any spirals but 14 did. These spirals were of
selected for
both sinistrorse and dextrorse types.
Taylor and Decker (1947), in a study of
143 isolates obtained from scabby potatoes,
beets, and radishes, the
criteria for their classification: acid-fastness;
starch hydrolysis; formation of dark brown
used following
145
surface ring on milk; acidification of milk;
reduction of nitrate to nitrite; utilization of
certain sugars, organic acids, and paraffin:
gelatin liquefaction; pigment formation
from tyrosine; and maximum growth tem-
perature. The only true correlation between
specific cultural properties and the ability to
produce typical lesions of potato scab was
obtained in the production of a dark brown
ring of surface growth on milk.
The following species may be tentatively
included in the Scabies series: S. scabies, S.
hawaviensis, and S. galtieri.
A number of other organisms isolated from
potato tubers or directly from the soil were
found capable of causing scab of potatoes
and must be included in this series. This is
true, for example, of A. violaceochromogenes
described by Krassilnikov (1949), and of A.
chromofuscus and A. prunicolor of Gause et
al. (1957). Other closely related forms have
been described, although pathogenicity tests
were not made.
A number of forms that apparently have
nothing to do with scab formation, but have
the characteristic properties of the series may
also be included.
XL. Series Lavendulae
Characteristic Properties
a. Sporophores straight or spiral-forming;
spores oval, smooth surface.
b. Aerial mycelium colored lavender to
pale blue.
c. Melanin-positive.
This is one of the true chromogenic series
of the genus Streptomyces. Organisms be-
longing to the Lavendulae series are widely
distributed in the soil and are represented
there by a large number of species and varie-
ties. Many of them are strongly antagonistic
and are capable of forming various important
antibiotics, such as streptothricin. Wood-
ruff and McDaniel (1958) reported that 90
per cent of all the antibiotics produced by
146
actinomycetes are streptothricins; here be-
long various related compounds, such as
streptin, streptolin, actinorubin, and anti-
biotic 136.
The most important species belonging to
this series are S. lavendulae and S. venezuelae,
organisms producing streptothricin and
chloramphenicol respectively.
S. lavendulae comprises organisms ex-
tremely variable in nature. Many of them
give rise, on cultivation, to different variants
or mutants. Some of these variants produce
a blue diffusible pigment on glucose-peptone
agar; others form a brown pigment. The sub-
strate mycelium of the blue pigment-forming
variants is pale blue, with scattered, small
pinpoint areas of deep blue. Upon complete
sporulation, the substrate growth becomes
covered with the characteristic lavender-
colored aerial mycelium; occasional sunken
areas have a slightly bluish tinge; the reverse
of the substrate growth is cream-colored
except for the small blue spots. Other vari-
ants produce a colorless to cream-colored
substrate growth free of any blue pigment
whatsoever; a brown diffusible pigment ap-
pears later, and the growth becomes covered
with thick lavender-colored mycelium. The
two types of variants are stable in nature.
Some variants may lose the capacity to
produce aerial mycelium.
S. venezuelae, as well, gives rise to a num-
ber of variants. Two strains were isolated
and found to be similar to S. lavendulae in
their cultural and physiological properties,
although they differed in their ability to
utilize various carbohydrates. Streptomyces
venezuelae utilizes arabinose, rhamnose,
xylose, lactose, and fructose; S. lavendulae
has either no effect or only a limited effect
upon these carbohydrates. The former also
differs from the latter in its sensitivity to
actinophage and in various serological re-
actions.
Streptomyces venezuelae was described as
having a thin-walled substrate mycelium,
colorless, hyaline, monopodially branched,
THE ACTINOMYCETES, Vol. II
the hyphae varying in diameter from 0.9 to
1.8 wand the branches growing to about 150
uw in length. The aerial mycelium appears
lavender under the microscope, thick-walled,
generally not much branched, straight or
shghtly and irregularly curved, not forming
spirals, individual hyphae arising frequently
from the primary mycelium at the surface
of the substrate. The color of the colonies,
when viewed on agar without magnification,
is gray to light tan or pink, but not lavender.
The upper portions of the aerial hyphae
divide into chains of spores. These are oval
to oblong 0.4 to 0.9 by 0.7 to 1.6 wu. Individ-
ual spores are colorless at maturity, but in
mass appear tan to gray when viewed with-
out magnification.
Okami (1956) made a comparative study
of the organisms commonly included in the
Lavendulae series on the basis of the color of
the aerial mycelum and certain other char-
acteristics. He found that eight cultures,
notably the streptothricin-producing forms,
possessed the following properties which he
considered as standard for the series.
a. Aerial mycelium pink-lavender color
when grown on yeast extract-glucose
agar.
b. Brown pigment when grown on yeast
extract-glucose agar.
c. Very sensitive to chlortetracycline and
chloramphenicol; relatively sensitive to
streptomycin; relatively resistant to
neomycin; and resistant to strepto-
thricin.
d. Utilize: glucose,
mannose. Do not utilize: arabinose,
fructose, lactose, mannitol, raffinose,
galactose, maltose,
rhamnose, sucrose, xylose.
e. They show certain growth-inhibiting
effects (Tables 24 and 25).
On continued cultivation for 40 years on
artificial media, the original 1915 isolate of
S. lavendulae (No. 3330) lost many of its
characteristic properties:
1. It no longer produced any aerial my-
celium.
SERIES AND SPECIES OF
2. It did not form any dark brown pig-
ment.
3. It was now able to utilize fructose,
mannitol, rhamnose, xylose.
4. It showed no or very weak antagonistic
action.
On the basis of the above properties,
Okami divided the S. lavendulae series into
10 subgroups (Table 26).
TABLE 24
Antibacterial activity of different strains of
Streptomyces lavendulae (Okami, 1956)
Sl lavendulae Inhibition zone, test bacteria
strain no. = E
B. cereus
y. coli Staph. aureus
mm mm mm
3330 0 0 0
3440-8 1t7/ 16 2
3440-14 6 6 2
3516 1S lg 2
3516-W 10 10 2
3530 16 16 2
3531 0 0 0
3532 27 23 18
3542 18 17 fel
3543 11 1s 6
3508 0) 0 i)
3445 0 0 0
3465 2 4 5
3544 11 i) 3
3559 21
GENUS STREPTOMYCES
considered S. lavendulae as a
Krassilnikov,
Baldacci
distinct group.
looked upon these organisms as members of
the ‘‘chromogenus” group. et al.
created a series under the name ‘‘/avendulae-
and subdivided it
groups, based upon the formation of a solu-
ble pigment in organic media: (1) The first
included cultures that formed
pigment, such as S. virginiae; (2) The second
included SS. venezuelae, S. lavendulae, and S.
circulatus, as well as a variety of others, all
of which formed a brown to black pigment
in organic media; (3) The third produced a
vellow pigment. As the requirements for the
group S. lavendulae presented above indicate,
neither the first nor the third of these sub-
groups belong to this group.
Sanchez-Marroquin (1958) found that
three related species of Streptomyces produc-
ing a pink to lavender aerial mycelium on
synthetic media could be distinguished from
one another as follows: (a) S. fradzae pro-
however,
(Gaause
roseus”’ into three sub-
no soluble
duces spirals on short branches; (b) S.
lavendulae forms spirals at the tip of long
straight branches; (c) S. venezuelae forms no
spirals, but only straight sporophores; the
first is nonchromogenic and the last two
are chromogenic. The Lavendulae series was
TABLE 25
Effect of composition of medium on reciprocal antagonism between different strains of
Streptomyces lavendulae (Okami, 1956)
Inhibition zone of S. lavendulae 3516
Inhibition zone of S. lavendulae 3440-8
Strain number Agar media*
Agar media
YGA == = - Soya YDA ———__—_—___—— Soya
ASP Starch ASP Starch
mm mm mm mm mm mm mm mm
3330 4.0 5.0 7.0 4.0 3.0 0 2.0 3.0
3440-8 17.0 15.0 20.0 15.0 0) i) 0) 0
3440-14 17.0 4.0 18.0 10.0 0 0 0 0
3516 0 0.5 0 0 0 0 0 0
3516-W 0 2.0 3.0 1.0 0 0 0 0
3530 0 1.0 1.0 1.0 0 0 0 0
3531 12.0 1.0 5.0 8.0 0 0 0 0
3532 16.0 10.0 2.0 12.0 7.0 0 10.0 10.0
* YGA = yeast extract-glucose; ASP = asparagine-glucose.
148
THE ACTINOMYCETES, Vol. II
TABLE 26
Classification of the lavendulae series into subgroups (Okami, 1956)
Subgroup Culture No. Characteristics
A Standard S. lavendulae
3440-8 a. Aerial mycelium with pink-lavender color on YDA
3440-14 b. Brown pigment on YDA
3516 c. Very sensitive to chlortetracycline and chloramphenicol, relatively
3516-W sensitive to streptomycin, relatively resistant to neomycin, resistant to
3530 streptothricin
3542 d. Utilizes: galactose, maltose, mannose
3544 Does not utilize arabinose, fructose, lactose, mannitol, raffinose, rham-
3555 nose, sucrose, xylose
e. Shows certain inhibiting effects
B 3330 a. No aerial mycelium
b. No dark brown pigment
d. Utilizes fructose, rhamnose, mannitol, xylose
e. Antagonistic action none or very weak
C 3531 c. Relatively resistant to chloramphenicol
D 3532 c. Relatively resistant to chlortetracycline
e. Antagonistic action none or weak
E 3543 b. No dark brown pigment
c. Relatively resistant to chloramphenicol
d. Utilizes arabinose, fructose, lactose, mannitol, raffinose, rhamnose, su-
crose, xylose
e. Antagonistic action none or weak
F 3568 d. Utilizes arabinose
e. Antagonistic action none or weak
G 3465 b. No dark brown pigment
e. Antagonistic action none or weak
H 3555 c. Relatively sensitive to streptothricin VI
d. Utilizes fructose
e. Antagonistic action none or weak
I 3625 c. Resistant to chloramphenicol
3534 d. Utilizes arabinose, rhamnose, xylose
3534-A e. Antagonistic action none or weak
J 3651* d. Does not utilize mannose
3652*
* S. virginiae
said to include not only S. lavendulae but also
S. cinnamonensis and SS. virginiae.
To indicate the confusion in classifying
of the series, It 1s
members Lavendulae
suficient to cite the work of Kuchayeva
(1958). She collected 22 cultures, freshly
isolated or obtained from different labora-
tories and believed to belong to this group.
—
SERIES AND SPECIES OF GENUS STREPTOMYCES
Fight of the cultures produced straight
sporophores and 14 formed spiral-shaped
sporophores. The color of the aerial myce-
lium of the substrate growth varied from
vellow to reddish-brown. Some produced a
melanin pigment and others did not, thus
automatically excluding the last as members
of the Lavendulae series. They varied greatly
in their antagonistic properties, some in-
hibiting the growth of all bacteria and fungi
tested, and others having no effect either on
gram-negative bacteria or on certain fungi.
The following antibiotics were listed as
products of the Lavendulae series, thus sug-
gesting the possible specific differences in
the series: streptothricin, streptin, antismeg-
matis factor, antibiotic 136, lavendulin,
actinorubin, pleocidin, ehrlichin, actithiazic
acid, antibiotic MD 2428, and grasseriomy-
cin.
XIL. Series Erythrochromogenes
This is a large melanin-positive series of
organisms. The aerial mycelium is usually
white with a yellowish or brownish shade to
gray with a bluish or greenish shade; it is
often reddish to The
growth is colorless to orange to red or even
brown to black. Certain species placed in
this group often produce a greenish-yellow
or reddish-brown to almost black soluble
brown. substrate
pigment on synthetic media. The sporo-
phores are straight or produce spirals.
This melanin-positive series comprises a
large number of species, such as represented
by the following: S. erythrochromogenes, S.
bobiliae, and S. cinereoruber.
XIIL. Series Viridochromogenes
This melanin-positive series of organisms
is characterized by an ash-gray to greenish
to olive-green to bluish aerial mycelium.
The substrate growth is cream-colored to
brown to greenish to black. Soluble pigment
on inorganic media is yellowish to greenish
149
to black. On organic media, soluble pigment
is brown to deep brown to olive or purple or
black.
The sporophores form spirals.
This series is widely represented in nature
by a number of species, namely: S. virido-
chromogenes, S. chartreusis, and S. cyaneus.
XIV. Series Cinnamomeus
Characteristic Properties
a. Sporophores produced in verticils.
b. Aerial mycelium white,
pinkish.
vellow, or
c. Substrate growth yellowish to pinkish.
d. Melanin-negative.
This series is characterized by the forma-
tion of verticil-bearing sporophores. The
verticils are both primary and secondary.
Spirals usually are not produced, although
occasionally some spirals are formed. The
species within this series are melanin-nega-
tive, although a purplish pigment may be
produced on certain organic media. The
aerial mycelium is white to pinkish to cin-
namon-colored. The substrate growth is
vellowish to brown to pinkish.
The species included in this series can be
listed here: S. hachijoensis, S. fervens, and
S. cinnamomeus.
XV. Series Reticuli
Characteristic Properties
I
a. Sporophores produced in verticils,
straight or spiral-shaped, on the primary or
secondary aerial hyphae.
b. Aerial mycelium white to gray.
c. Melanin-positive.
This is one of the two series within the
genus Streptomyces which are differentiated
from the other members of the genus pri-
marily by their morphology. Species within
this series are characterized by the radial
arrangement of the sporophores, whereby
three or more branches originate from a
150
node, thus forming a ‘‘verticil,”’ frequently
referred to as a ‘‘whorl.’? Very often the
sporophores may be branched toward the
end of the sporulating-hyphae, giving the
appearance of a “broom shaped branch,”’
or may give rise to the formation of a
“cluster,” which is to be distinguished from
a typical verticil of sporophores. The verti-
cils may be primary or secondary in nature.
Among the variations of a verticil is the
formation of tufts, when straight branches
are grouped together on the aerial hyphae.
The formation and nature of the verticils
may be changed with the composition of the
medium, a phenomenon first reported by
Waksman and Curtis (1916) and more
recently by Nakazawa (1955) and Shinobu
(1955a). Both primary and secondary ver-
ticils may be formed in the same culture.
Baldacci (1953) did not recognize this im-
portant series at all. He gave series recog-
nition to one of its members, S. rubrireticult,
merely on the basis of its pigmentation.
Gause et al. (1957), following this example,
included in the ‘ruber’ series one ‘‘tuft-
forming” organism, also on the basis of its
pigmentation. Later, however, Baldacci
(1958) suggested separation of the verticil-
producing organisms into a separate genus,
Streptoverticillium, as
(Chapter 4, Volume I).
Various investigators have used morphol-
shown — elsewhere
ogy as a basis for series separation of the
genus Streptomyces. It is sufficient to list
here three of them.
Shinobu (1955a) proposed division of the
genus as follows:
1. Monopodial branching. This section has
been divided on the basis of spiral forma-
tion.
2. Verticil formation:
a. Nitella type. Typical radial branches
almost equal in length. No spirals
formed. S. reticulz is given as a typical
representative.
b. Anitella type. Radial branches differ
THE ACTINOMYCETES, Vol. II
from each other. Spirals formed with-
out proper radial symmetry. S. virido-
chromogenes is representative.
The S. reticuli subgroup was further di-
vided into:
A. Verticils only primary. Sporophores straight.
Streptomyces verticillatus
Streptomyces hiroshimensis
B. Verticils both primary and secondary. Sporo-
phores form spirals.
I. Colorless to brownish growth on synthetie
media.
1. Good growth on protein media. Spores
spherical, oval.
Streptomyces reticult
Streptomyces albireticuli
2. Poor growth on protein media. Spores
cylindrical.
Streptomyces circulatus
II. Growth on synthetic media pink to red.
Streptomyces reticuloruber
Streptomyces griseocarneus
Ill. Growth on synthetic media greenish.
Streptomyces verticilloviridans
Solovieva et al. (1957) made a study of
cultures belonging to the S. reticuli series
and isolated from Pamir soils. These cultures
were divided into two subgroups:
I. S. verticillatus, with straight sporophores
(primary and secondary).
Il. S. reticulz, with spiral-shaped sporo-
phores.
These subgroups showed very little dif-
ference in their physiological and biochem-
ical properties. Only one strain, S. rubri-
reticuli, showed some differences (weaker
gelatin liquefaction, strong nitrate reduction,
weak growth on cellulose). From an anti-
biotic point of view, however, there was a
marked difference; members of the first sub-
group showed strong antifungal activity,
whereas subgroup II gave weaker activity
or none.
Pridham et al. (1958) divided the verticil-
forming series into four morphological sub-
groups:
1. Monoverticillate, no spirals.
2. Monoverticillate, with spirals.
SERIES AND SPECIES OF
3. Biverticillate, no spirals.
4. Biverticillate, with spirals.
The following species were included in this
series: S. reticuli, S. netropsis, S. thioluteus,
S. griseocarneus, and S. verticillatus.
Unfortunately, some cultures show both
straight and — spiral-shaped
Nomi (1960) criticized the above system;
he could not accept the spiral-producing
forms among the true verticil types. He con-
sidered the group to consist of the typical
sporophores.
“biverticils’”” comprising both primary and
secondary elements; the atypical ‘‘mono-
verticils’ comprising primary — verticils,
mixed verticils, and sometimes more com-
pound polyverticils. He included the follow-
ing species: S. reticult, S. hiroshimensis, S.
albireticuli, S. echimensis, S. griseocarneus,
S. thioluteus, S. salmonicida, and 8. netropsis.
GENUS STREPTOMYCES 15]
Series Thermophilus is discussed in Chap-
ter 11, among the thermophilic actino-
mycetes.
Other Possible Series
In addition to these 16 series of the genus
Streptomyces, other series could be suggested,
based either on the color of the aerial myce-
lium or of the substrate growth; ecological
or physiological criteria have also been pro-
posed for series characterization. Certain
groups, such as the thermophilic forms be-
longing to the genus Streptomyces are in-
cluded in a special series (Vhermophilus),
and are discussed elsewhere (Chapter 10),
since they are definitely related, because of
their close ecological and physiological re-
lationships, to the other thermophilic genera.
Cohva'p teu, 77
Classification of Streptomyces Species
The difficulties encountered by an inex-
perienced investigator in identifying a
freshly isolated culture of a Streptomyces with
previously described species have been
brought out in the preceding three chapters.
No wonder an inexperienced worker soon
becomes discouraged and takes the easy path
of creating a new species or variety for
every newly isolated culture. This is es-
pecially true if such a culture produces an
antibiotic not previously described or an
apparently different form of a known anti-
biotic that he is anxious to patent or on
which he wishes to establish priority. Among
the significant factors that have contributed
to this rash of “new” species are:
1. Inadequate description of previously
described species with which comparisons
are made.
2. Overlapping of the morphological and
cultural characteristics of strains or species
previously described.
3. Variations in composition of the media
used in describing species.
4. Failure to recognize natural variability
of different strains that might be meluded
in a single species.
5. Idiosynerasies of the particular inves-
tigator, and his tendency to be either a
“Jumper” or a “‘splitter.”’
In spite of these discouraging aspects of
the problem of identification of particular
organisms, Classification and characteriza-
tion of Streptomyces species have recently
made considerable progress. Though various
criticisms have been directed toward it, the
system of classification of actinomycetes
used in the seven editions of Bergey’s Man-
ual still appears to be the most logical and
most workable, except for certain modifica-
tions that are now desirable. In every new
edition, advantage has been taken of the
accumulated information to modify this sys-
tem of classification, especially of the genus
Streptomyces. In this treatise, a complete re-
arrangement has been made in classifying
the species included in the genus Strepto-
myces, as compared with the last (seventh)
edition of Bergey’s Manual.
The thermophilic species of Streptomyces
have been placed in a separate series Thermo-
philus and transferred to Chapter 11, in
which all the thermophilic actinomycetes
are included. Species of Streptomyces 1so-
lated from animal and plant infections, es-
pecially those for which pathogenicity has
not been established, have been distributed
throughout the genus, thus removing the
need for a major separation of these species
into saprophytic versus parasitic forms.
These properties are now given only sec-
ondary consideration in characterizing spe-
cies within the genus. It has further been
recognized and emphasized, time and again,
that it is most desirable to utilize morpho-
logical properties in defining and charac-
terizing species of Streptomyces, though in
some instances this could not be done with
any degree of assurance.
New genera have been created, once cer-
tain well-defined morphological and physio-
logical properties suggested its advisability.
:
CLASSIFICATION OF STREPTOMYCES SPECIES
Whether this practice should be extended
and recognition thus be given to the capacity
to form sclerotia or to the ability to form
verticils by certain Streptomyces species,
thus placing them in separate genera, re-
mains to be determined. The author’s sug-
gestion of many years ago that the structure
of the sporophores (straight versus spiral-
forming, closed versus open spirals, tuft- and
verticil-producing) be used in characterizing
certain species or species-groups is gaining
wider recognition, though some investiga-
tors do not consider this a sufficiently con-
stant property for the major subdivision of
the genus and suggest that it be left for
secondary characterization. The shape and
size of the spores appear to be less signifi-
‘ant properties, although the surface of the
spores, as detected by the electron micro-
scope, has been gaining approval.
Among the most annoying characteristics
of the genus Streptomyces are: (a) the loss of
capacity by certain species to produce aerial
mycelium, and (b) the overlapping property
on the part of certain species of Nocardia to
produce an aerial mycelium that cannot be
differentiated from that of Streptomyces. It is
true, however, that cultures of Streptomyces,
even if they have lost the capacity to pro-
duce aerial mycelium, can still be recognized
by the structure of their substrate mycelium
and by certain cultural and physiological
properties. The latter include the nature of
their soluble pigments, their ability to
liquefy gelatin, hydrolyze starch, invert
sucrose, and coagulate and peptonize milk.
Some strains that have lost the capacity to
produce aerial mycelium may regain this
property if they are grown in sterile soil or
in special soil media, or are subjected to
other special treatments.
The growth of cultures of Streptomyces
that have lost the capacity to produce aerial
mycelium is often colorless, though some-
times pigmented; it is smooth or lichenoid,
leathery, compact, with a shiny surface.
Some produce a soluble brown pigment.
Some are able to form antibiotics. On the
assumption that such cultures, because they
do not form aerial spores, should be con-
sidered as sterile, Krassilnikov designates
them as trmomials with a third component
of the name “‘sterilis.’”’ This is analogous to
Fungus sterilis among the fungi. Certain
such species are included in the present
classification; others may be considered as
typical nocardias and have been transferred
to that genus.
Characterization of Streptomyces Spe-
cies
Among the properties to receive major
consideration in describing individual spe-
cies are the following:
1. Morphological properties: These include
formation and nature of substrate (vegeta-
tive) growth and of aerial mycelium, manner
of sporulation (spiral formation, verticil
formation), nature and surface of spores.
2. Cultural characteristics. These
prise color and color changes of substrate
growth and of aerial mycelium, and forma-
tion of soluble pigments on synthetie and
com-
organic media. The most significant of these
pigments are the melanins or melanoids pro-
duced in media containing tyrosine or pro-
teins and peptones. The brown to black
pigments produced in such media by certain
species of Streptomyces are said to designate
melanin-positive as opposed to melanin-
negative reactions, or tyrosinase-positive
versus tyrosinase-negative reactions. In view
of the fact, however, that few of the older
investigators tested this reaction in tyrosine—
free media, it is more desirable to use the
designations chromogenic (melanin +) or
nonchromogenic (melanin —). In this case,
chromogenicity the
formation of brown to black pigments on
refers specifically to
protein-containing media.
3. Physiological and biochemical properties.
These include: proteolytic activities, such as
154
gelatin liquefaction, coagulation and pep-
tonization of milk, and hemolysis of blood;
utilization of carbon compounds; antagonis-
tic properties and formation of antibiotics;
effect of temperature, aeration, and reaction
upon growth; formation of specific enzymes,
such as oxidase, lipase, invertase, diastase,
mannase and protease; reduction of nitrate,
and formation of H.S. There is no sharp line
of demarcation between cultural and physio-
logical properties, on the one hand, and be-
tween physiological and biochemical activi-
ties, on the other.
4. Ecology. The ability of the organism to
cause animal or plant diseases, and its occur-
rence in a natural environment are important
characteristics.
5. Supplementary characteristics. Addi-
tional characteristics that may be utilized
for descriptive purposes include: (a) sero-
logical reactions, (b) phage sensitivity, and
(c) sensitivity to specific antibiotics.
In describing the various species, one has
to depend frequently upon the mformation
supplied in published reports, since type
cultures often are not available. Where the
desired information is lacking or where the
description, for various reasons, Is iInade-
quate, the species may be placed in the list
of incompletely described forms.
tecently there has been a tendency to
overlook earlier described species, and to
emphasize and often give new names to
newly isolated cultures. One cannot con-
demn this tendency too strongly. While, m
most cases, it is dificult to establish synon-
vmy because of a lack of type cultures or a
possible change in such cultures upon pro-
longed cultivation on artificial media, every
effort must be made to give credit to the
sarlier investigator. One has no patience,
therefore, with those attempts to set aside,
willfully or unwillfully, older descriptions
or to consider such organisms as varieties of
newly isolated and newly named cultures.
When a culture is freshly isolated, a study
THE ACTINOMYCETES, Vol. II
should be made both of its position in a
particular group in the genus and of its
classification as a species. Only by a com-
bination of properties described under both
can one determine the identity of the new
culture. Obviously, no conclusions should be
drawn that such a culture represents a new
species merely on the basis of certain super-
ficial observations, such as a delay in coag-
ulation and peptonization of milk or in
liquefaction of gelatin, or because of a differ-
ence in the intensity of coloration of the
potato plug, or in the shade of pigmentation
of growth on a particular medium, or even
in the degree of curvature of the sporophores.
Any attempt to create new species on the
basis of such minor variations must be con-
sidered both unscientific and confusing. The
tests must be repeated again and again to
confirm the recorded observations.
The need for a knowledge of the exact
composition of the media used for descrip-
tive purposes can hardly be overemphasized.
Some species show their most characteristic
property upon only one particular medium,
and unless such a medium is used, the prop-
erties of a new isolate can easily be over-
looked. S. fradiae, for example, shows the
characteristic color (seashell-pink) of its
aerial mycelium upon potato-starch agar.
Certain synthetic agar media, notably
nitrate-sucrose, glucose-asparagine, and cal-
cium malate, are among those which alone
bring out characteristic properties of certain
other species. Growth on potato is quite
characteristic, although the variety of the
potato, the manner of crop fertilization, and
other factors may influence the nature of the
growth of the organism and pigment formed.
In view of the great interest at the present
time in the screening programs for antibi-
oties, when literally many thousands of cul-
tures are being isolated and tested, there is
naturally a tendency on the part of some
investigators to consider the ability of a
particular culture to form a specific anti-
CLASSIFICATION OF STREPTOMYCES SPECIES
biotic as its major characteristic property.
But this capacity is often a strain rather than
a species characteristic. In view of the mu-
tational possibilities of such cultures and of
the marked effect of composition of medium
and environmental factors upon the qualita-
tive nature and quantitative yield of the
antibiotic, one must consider such a prop-
erty, at best, as only a secondary character-
istic and avoid assigning to it an important
role in creating new species.
Shinobu (1958b) emphasized again that
only synthetic media should be used for the
study of the sporulation of the aerial myce-
lium, notably spiral formation, in Strepto-
myces. In most species, the curvature of the
spiral is sinistrorse (counterclockwise); In a
few, dextrorse (clockwise). The diameter of
the spiral varies from 1.5 to 8.0 uw and is not
a characteristic property, although it is fixed
for some species. As pointed out previously
(Chapter 4), three morphological groups
were recognized: (1) Those forming straight
or wavy aerial mycelium, (2) Spiral-forming
types, and (3) Verticil-forming types.
Other criteria have been suggested. Some
of these may be utilized for supplementary
information in describing species and vari-
eties. A Subcommittee on Actinomycetes of
the Society of American Bacteriologists
(Gottlieb, 1960) gave careful consideration
to the various criteria used in describing and
characterizing species of Streptomyces. They
‘ame to the conclusion that morphology is
to be considered as one of the more impor-
tant criteria. Color of the aerial mycelium
and carbon utilization are also important.
Supplementary characteristics are provided
by the production of HS, reduction of
nitrate, and gelatin liquefaction. Such cri-
teria as color of substrate growth and nitro-
gen utilization were not considered of suffi-
cient significance in describing new species.
Many species, including a number of
newly isolated forms, have been placed in
Chapter 13 as incompletely described. Others
—
On
Or
have been listed as synonyms. Care has been
taken to avoid the creation of many
varieties, unless it has been fully established
new
that such varieties have a sound morpho-
logical or cultural Whether
varieties should be raised to the status of
species remains to be determined by further
study. Any such attempt would automat-
ically lead to the temptation to create new
species out of mutants, which are unfortu-
nately altogether whether
naturally occurring or artificially created.
For the time being, they may still be con-
sidered as varieties.
basis. these
too common,
Classification of Genus Streptomyces
In proposing the present system of clas-
sification of the genus Streptomyces, the
following properties have been given the
greatest consideration:
lL. Morphology of sporulating bodies; size,
shape, and surface of spores.
2. Color of aerial mycelium and of sub-
strate growth.
3. Color of growth.
4. Formation of soluble chromogenic or
melanoid pigments in proteinaceous media.
This property is used, together with micro-
morphology, for the major subdivisions of
the genus.
5. Formation of soluble pigments in syn-
thetic media.
6. Certain biochemical properties, notably
proteolysis, starch hydrolysis, nitrate reduc-
tion, formation of HS, utilization of carbon
sources, and formation of specific anti-
biotics.
This system is a modification of the one
originally used by Waksman and Curtis in
1916, variously changed in subsequent years
by Waksman and by Jensen, and used in
the various editions of
Bac-
modified forms in
Bergey’s Manual of Determinative
teriology.
A. Sporophores straight, wavy, or spiral-shaped;
no verticils.
156 THE ACTINOMYCETES,
I. Proteinaceous mediaare not pigmented deep
brown or black; melanin—negative. Soluble
pigment on various media is faint brown,
pink, red, purple, yellow, blue, or absent.
1. Soluble pigment only faint yellow or
faint brown.
a. Aerial mycelium white, abundant.
al. Sporophores produce spirals.
a®. Occurs in soil and in certain
other natural substrates.
15. Streptomyces albus
b?. Occurs in the sea.
148. Streptomyces marinus
b!. Sporophores straight.
a2, Sporophores produce broom-
shaped clusters.
102. Streptomyces globispo-
rus
b2. No clusters produced.
a*. Nonproteolytic.
27. Streptomyces autotro-
phicus
b’. Weakly proteolytic.
179. Streptomyces orientalis
ec’. Strongly proteolytic.
129. Streptomyces kimbert
b. Aerial mycelium, white, scant.
250. Streptomyces willmoret
c. Aerial mycelium white to light gray.
al. Sporophores produce compact
spirals.
a®. Growth colorless.
18. Streptomyces annula-
tus
—
~
[<)
2, Growth colorless to yellow-
ish.
92. Streptomyces fungict-
dicus
io)
t
Growth brown to. orange-
brown.
21. Streptomyces arenae
d?. No growth on sucrose nitrate
agar.
22. Streptomyces argenteo-
lus
b!. Sporophores form loose spirals.
40. Streptomyces calvus
c!. Sporophores straight or wavy.
a®. Growth on sucrose nitrate
agar yellowish-brown.
2. Streptomyces — abura-
vieNnsts
b?. Growth gray to greenish.
60. Streptomyces — coroni-
formis
Vol-vil
e2. Growth vellowish-white;
utilizes paraffin and rubber.
68. Streptomyces elasticus
. Aerial mycelium white to mouse-
gray; spores bluish-gray.
al. Sporophores produce compact
spirals.
227. Streptomyces spheroi-
des
b!. Sporophores in clusters; a few
compact spirals.
45. Streptomyces catenulae
c!. Sporophores produce open cork-
screw spirals.
a®. Soluble pigment yellow.
a’. Growth on — synthetic
media cream-colored.
167. Streptomyces niveus
b*. Growth on synthetic me-
dia vellow.
143. Streptomyces — macro-
sporeus
b?. Soluble pigment tan to
brown.
35. Streptomyces caelestis
!| Sporophores straight.
111. Streptomyces griseolus
Aerial mycelium white to gray,
covered with dark humid stains or
guttation drops.
a'. Sporophores form spirals.
a®. Growth buff to olive-colored.
189. Streptomyces platensis
b?. Growth colorless.
123. Streptomyces humidus
b!. Sporophores straight.
a®. Growth on potato cream-
colored.
+
pou
61. Streptomyces craterifer
b?. Growth on potato slimy to
black.
235. Streptomyces tumult
Aerial mycelium green.
al. Growth green.
196. Streptomyces prasinus
b!. Growth colorless.
121. Streptomyces hirsutus
c!. Growth red.
195. Streptomyces prasino-
pilosus
Aerial mycelium limited, produced
late; white with tinge of gray.
97. Streptomyces gardnert
Soluble pigment blue or purple.
CLASSIFICATION OF STREPTOMYCES SPECIES 157
a. Aerial mycelium white.
63. Streptomyces cyanofla-
pus
b. Aerial mycelium white to gray.
al. Soluble pigment produced only
on potato and certain other
media; pigment changes to red
in an acid and to green in an
alkaline reaction.
58. Streptomyces coelicolor
b!. Pigment produced on all media,
red in an acid and blue in an
alkaline reaction.
240. Streptomyces violaceo-
ruber
c!, Pigment at first yellow-red,
changing to blue or bluish-green.
190. Streptomyces plurt-
color
d'. Pigment purple.
171. Streptomyces novaecae-
sareae
e!. Soluble pigment bluish to black.
239. Streptomyces violaceo-
niger
e. Aerial mycelium blue.
al. No spirals formed.
36. Streptomyces caeruleus
bt. Open spirals.
241. Streptomyces violaceus
3. Pigment at first green, becoming
brown.
a. Aerial mycelium usually absent.
237. Streptomyces verne
b. Aerial mycelium white.
221. Streptomyces samp-
sonit
ce. Aerial mycelium brownish-white to
brownish-gray.
187. Streptomyces phaeovi-
ridis
d. Aerial mycelium dark gray, olive-
colored, or grayish-green; sporo-
phores produce spirals.
244. Streptomyces viridans
4. Pigment yellow to golden yellow.
a. Growth green to greenish-yellow.
al. Aerial mycelium weakly de-
veloped; white or pale yellow.
242. Streptomyces virgatus
b!. Aerial mycelium gray to dark
gray.
125. Streptomyces interme-
dius
c!. Aerial mycelium scant, white.
Cc.
d.
e.
144. Streptomyces macula-
tus
d'. Aerial mycelium — cinnamon-
colored.
158. Streptomyces murinus
. Growth green to dark green.
14. Streptomyces alboviri-
dis
Growth sulfur-yellow.
at. Aerial mycelium white to pink-
ish.
81. Streptomyces flaveolus
bt. Aerial mycelium light yellow.
182. Streptomyces parvus
c!. Aerial mycelium white to gray
to reddish-gray.
251. Streptomyces xantho-
phaeus
d'. Aerial mycelium light gray.
48. Streptomyces cellulosae
e'. Aerial mycelium ash-gray.
181. Streptomyces parvullus
f'. Aerial mycelium yellowish-green
to sulfur-yellow.
230. Streptomyces sulphu-
reus
Growth carmine red, reddish-brown
to orange-colored to cinnamon-drab.
al. Aerial mycelium chalk-white.
166. Streptomyces niveoru-
ber
b!. Aerial mycelium white to gray.
a®. No aerial mycelium on _ po-
tato.
208. Streptomyces rimosus
b?. Aerial mycelium on potato
white to gray to black.
25. Streptomyces aureofa-
clens
Closely related form.
222. Streptomyces saya-
Maensis
c?. Aerial mycelium on potato
olive-buff.
57. Streptomyces clavifer
c!. Aerial mycelium grayish-brown.
220. Streptomyces saha-
chirot
d'. Aerial mycelium yellowish-gray.
110. Streptomyces griseofla-
vus
Growth cream-colored to brown.
al. Rapid liquefaction of gelatin.
7. Streptomyces albidofla-
vUuS
158 THE ACTINOMYCE?
bt. Gelatin slowly liquefied.
50. Streptomyces chibaen-
sis
5. Soluble pigment yellowish to yellow-
green.
a. Aerial mycelium white.
51. Streptomyces
mallus
b. Aerial mycelium white to yellow.
al. Growth yellowish to green.
134. Streptomyces liesker
bt. Growth yellow, becoming black.
135. Streptomyces limosus
ce. Aerial mycelium gray.
al. Growth on sucrose nitrate agar
yellowish-green.
85. Streptomyces — flavovi-
chryso-
rens
b!. Growth on sucrose nitrate agar
yellow.
47. Streptomyces — cellulo-
flavus
6. Soluble pigment yellowish-brown to
reddish-brown.
a. Growth cream-colored.
al. Sporophores flexible and hooked.
112. Streptomyces griseolu-
teus
b!. Sporophores produced in clus-
ters.
165. Streptomyces nitrospo-
reus
b. Growth has rosy tinge.
204. Streptomyces ramnait
ce. Growth yellowish.
17. Streptomyces ambofa-
clens
d. Growth has reddish tone.
191. Streptomyces pluricolor-
escens
e. Growth becoming red.
al. Aerial mycelium white.
70. Streptomyces eryth-
raeus
b'. Aerial mycelium white to gray
with greenish tinge.
205. Streptomyces ramulo-
sus
c'. Aerial mycelium mouse-gray to
drab.
83. Streptomyces flavogri-
seus
d'. Aerial mycelium white to gray
to olive-buff.
224. Streptomyces setonii
oe)
TES, Vol. II
Soluble pigment in synthetic media
brown.
a. Growth coral-red.
24. Streptomyces aurantia-
cus
b. Growth yellow.
32. Streptomyces
pensis
c. Growth yellowish-brown
3. Streptomyces achromo-
genes
d. Growth brown to purplish.
16. Streptomyces althioti-
cus
e. Growth black.
162. Streptomyces niger.
Soluble pigment on potato plug brown
to brownish-red to reddish-purple.
a. Growth on potato greenish-colored.
al. Spirals formed.
65. Streptomyces diastati-
cus
b!. No spirals.
142. Streptomyces lydicus
b. Growth on potato gray; no spirals
formed.
bottro-
42. Streptomyces canescus
c. Growth on potato yellowish-colored.
al. Aerial mycelium white.
87. Streptomyces flocculus
b!. Aerial mycelium gray to yellow-
ish.
a®. Growth cream-colored.
80. Streptomyces — fimica-
rlUus
b?. Growth yellow-brown.
75. Streptomyces felleus
d. Growth on potato yellow turning
white.
147. Streptomyces marinoli-
MOSUS
e. Growth on potato pink to reddish-
purple.
al. Sporophores produce spirals.
a2. Aerial mycelium cinnamon
to drab-gray.
170. Streptomyces nourser
b?. Aerial mycelium gray.
11. Streptomyces albogri-
seolus
b!. Sporophores both straight and
spiral-forming.
228. Streptomyces spiralis
c!. Growth on various media yellow-
orange to brown.
89. Streptomyces fragilis
oP
10.
CLASSIFICATION OF STREPTOMYCES SPECIES 159
Soluble pigment on synthetic agar
brown to black.
a. Growth on potato gray to brown.
74. Streptomyces exfolia-
lis
b. Growth on potato greenish to black.
100. Streptomyces gelaticus
c. Aerial mycelium pigmented green.
101. Streptomyces glaucus
No soluble pigment on synthetic media.
a. Growth yellowish-brown.
160. Streptomyces narbo-
nensis
b. Growth yellowish to pink to black.
al. Aerial mycelium abundant, gray.
23. Streptomyces armilla-
tus
bl. Aerial mycelium white, with
pinkish to orange tinge on cer-
tain media.
77. Streptomyces filamen-
tosus
e'. Aerial mycelium white to yellow.
127. Streptomyces kanamy-
celicus
d'. Aerial mycelium white to pink.
145. Streptomyces madurae
e!. Aerial mycelium scant, white.
1838. Streptomyces pelletiert
f'. Aerial mycelium white-gray to
black.
225. Streptomyces somalien-
sis
' Aerial mycelium black.
180. Streptomyces paraguay-
ge
ensts
ce. Growth yellowish to orange.
al. Aerial mycelium white to rose-
colored.
213. Streptomyces roseofla-
vus
b!. Aerial mycelium white.
201. Streptomyces putrificus
c!. Aerial mycelium scant, white to
grayish-brown.
90. Streptomyces fulvisst-
mus
d'. Aerial mycelium orange to pale
pink.
226. Streptomyces spectabi-
lis
e!, Aerial mycelium yellowish to
gray.
86. Streptomyces flavus
f!. Aerial mycelium has olive tinge.
122. Streptomyces hominis
d.
g!. Aerial mycelium scant, rose-
vellow.
155. Streptomyces microfla-
vUS
h!. Aerial mycelium white to orange-
colored.
216. Streptomyces ruber
i'. Aerial mycelium gray to mouse-
gray.
34. Streptomyces cacaoi
Growth yellowish-green to citron-
yellow; aerial mycelium white to
yellow to pinkish.
56. Streptomyces citreus
Growth colorless to cream-colored.
al. Aerial mycelium scant, white.
a?. Good growth on milk.
104. Streptomyces gougero-
tia
b?. No growth on milk.
113. Streptomyces — griseo-
planus
b!. Aerial mycelium white to olive-
buff.
193. Streptomyces praecox
e!. Aerial mycelium white.
a®. Acid-sensitive.
8. Streptomyces albidus
b?. Acid-resistant.
5. Streptomyces acidophi-
lus
d'. Aerial mycelium white to gray.
a®. Sporophores straight.
249. Streptomyces wedmor
ensis
b?. Sporophores produce spirals.
197. Streptomyces pseudo
griseolus
e'. Aerial mycelium sandy lavender
to dark gray.
209. Streptomyces rochet
f'. Aerial mycelium rose-colored.
215. Streptomyces roseus
Growth black.
99. Streptomyces gedanen-
sts
Growth yellow to olive-ocher.
174. Streptomyces olivaceus
. Growth colorless to yellowish to
olive-buff. Aerial mycelium water-
green.
al. Green and yellow pigments on
malate and succinate media.
116. Streptomyces griseus
160 THE ACTINOMYCETES, Vol. II
b!. No green and yellow pigments
on malate and succinate media.
106. Streptomyces griseinus
i. Growth red or purple.
39. Streptomyces californi-
CUS
j. Growth colorless to black.
al. Aerial mycelium white to brown-
ish-gray.
202. Streptomyces pyrido-
myceticus
b!. Aerial mycelium on synthetic
media dull gray.
157. Streptomyces mita-
kaensis
k. Growth dark brown.
al. Sporophores produce spirals.
a2. Aerial mycelium white to
gray.
a®. Growth on potato has
green tinge.
118. Streptomyces halstedii
b*. No green tinge on potato.
219. Streptomyces rutgers-
ensts
b?. Aerial mycelium olive-gray.
169. Streptomyces nodosus
b!. Sporophores straight.
a2, Aerial mycelium gray-white.
91. Streptomyces fumosus
b?. Aerial mycelium dark gray.
136. Streptomyces lipmanti
1. Growth on synthetic media rose to
gray.
212. Streptomyces roseodia-
staticus
m. Growth cream-colored to yellow or
vellow-orange.
al. Aerial mycelium on certain me-
dia white, moist with dark,
glistening patches.
124. Streptomyces hygrosco-
picus
bt. Aerial mycelium white-yellow
to brownish-yellow.
138. Streptomyces — longis-
poroflavus
c'. Aerial mycelium white.
a®. Aerial mycelium present on
protein media.
41. Streptomyces candidus
b?. Aerial mycelium absent on
protein media.
178. Streptomyces omiyaen-
Svs
B:
q.
d'. Aerial mycelium powdery white,
with vellow tinge.
a®. Little spiral formation.
10. Streptomyces albofla-
vus
. Abundant spiral formation.
43. Streptomyces canus
e!. Aerial mycelium gray.
20. Streptomyces antimy-
colicus
b2
~
t
. Growth colorless to pinkish to
brown.
159. Streptomyces naga-
nishit
Growth orange or red.
al. Growth yellowish to orange;
aerial mycelium seashell-pink.
a®. Produces antibacterial (neo-
mycin) and antifungal (fra-
dicin) antibiotics.
88. Streptomyces fradiae
b?. Produces antiviral (luridin)
agent.
140. Streptomyces luridus
b!. Growth rose to red; aerial my-
celium white.
a2. Growth yellow to red; weak
proteolysis.
13. Streptomyces albospor-
eus
2. Growth pale pinkish-buff;
strong proteolysis.
33. Streptomyces brasilien-
Sis
c!. Growth pale rose to red; aerial
mycelium weakly developed,
velvety, rose-white.
173. Streptomyces otdio-
sporus
d'. Growth red; aerial mycelium
black.
152. Streptomyces melano-
~
b
cyclus
Growth colorless, turning dark.
69. Streptomyces endus
Growth beeoming — salmon-pink;
acid-sensitive.
217. Streptomyces rubescens
Growth green to dark green; aerial
mycelium whitish to grayish.
245. Streptomyces viridis
Growth on blood agar brick-red.
164. Streptomyces — nitrifi-
cans
CLASSIFICATION OF STREPTOMYCES SPECIES 16]
II. Proteinaceous media are pigmented deep
brown to black; melanin-positive.
1. Growth colorless on synthetic media.
a. Aerial mycelium thin, rose-colored.
al. Spirals produced.
210. Streptomyces roseo-
chromogenes
bt. No spirals formed.
54. Streptomyces cinnamon-
ensis
b. Aerial mycelium white with pale
pink or pale gray tinge.
131. Streptomyces — kitasa-
waensis
ec. Aerial mycelium gray to brown to
reddish.
al. Growth on organic media green-
ish to black.
175. Streptomyces olivochro-
mogenes
bt. Growth dark brown.
206. Streptomyces — resisto-
myctficus
c!. Growth cream-colored.
163. Streptomyces nigrifa-
clens
d. Aerial mycelium cottony, dark
brown.
66. Streptomyces diastato-
chromogenes
e. Aerial mycelium pale yellow to gray.
30. Streptomyces blastmy-
ceticus
bho
7~
‘owth on synthetic media yellow.
a. Aerial mycelium white.
203. Streptomyces rameus
b. Aerial mycelium white to gray.
82. Streptomyces flavochro-
mogenes
c. Aerial mycelium white to yellow.
46. Streptomyces cavouren-
SIS
d. Aerial mycelium ash-gray.
188. Streptomyces pilosus
e. Aerial mycelium mouse-gray to
green-gray.
94. Streptomyces galbus
f. Aerial mycelium hazel brown.
139. Streptomyces lucensis
g. Aerial mycelium olive-buff.
al. Soluble pigment green to olive
to black.
233. Streptomyces tenuis
bt. Soluble pigment cream-colored
to golden brown.
146. Streptomyces margina-
tus
h. Aerial mycelium white with patches
of bluish-green on starch media.
126. Streptomyces ipomoeae
3. Growth white to gray.
a. Sporophores produce spirals.
al. Causes potato scab.
223. Streptomyces scabies
b!. Does not cause potato scab.
a’, Growth on potato gray.
119. Streptomyces hawaii-
ensis
b?. Growth on potato orange-
red.
96. Streptomyces galtieri
b. Sporophores straight.
at. Aerial mycelium white to gray.
29. Streptomyces bikinien-
Sis
bt. Aerial mycelium white, cottony.
156. Streptomyces mirabilis
c. Sporophores tend to be straight;
spirals less marked.
28. Streptomyces beddardii
4. Growth cream- to brown-colored.
a. Sporophores in clusters.
al. Aerial mycelium on _ nutrient
agar gray to yellowish-green.
19. Streptomyces antibioti-
Cus
bt. Aerial mycelium on nutrient
agar ash-gray.
73. Streptomyces euryther-
mus
c!. Aerial mycelium on nutrient
agar white.
38. Streptomyces caiusiae
b. Sporophores not in clusters.
al. Aerial mycelium white to gray.
109. Streptomyces — griseo-
chromogenes
b!. Aerial mycelium olive-gray.
248. Streptomyces _ virido-
genes
c!. Aerial mycelium olive-buff to
water-green.
107. Streptomyces — griseo-
brunneus
5. Growth red to reddish-orange.
a. Aerial mycelium white.
200. Streptomyces purpur-
ascens
162 THE ACTINOMYCETES, Vol. II
b. Aerial mycelium white to gray.
114. Streptomyces
ruber
griseo-
ce. Aerial mycelium gray.
al. No soluble pigment on synthetic
media.
95. Streptomyces galilaeus
b!. Soluble pigment on synthetic
media light carmine.
52. Streptomyces cinereo-
ruber
d. Aerial mycelium secant; ability to
produce such mycelium easily lost.
31. Streptomyces bobiliae
e. Aerial mycelium pink, with bluish-
green spores.
28a. Streptomyces bellus
6. Growth white to cream-colored.
98. Streptomyces garypha-
lus
Growth buff to dark brown.
a. Aerial mycelium gray to dark olive.
105. Streptomyces gracilis
b. Aerial mycelium dark gray.
103. Streptomyces globosus
ce. Aerial mycelium white.
64. Streptomyces cylindro-
~I
Sporus
d. Aerial mycelium tan to light brown.
115. Streptomyces griseovi-
ridis
Growth on synthetic agar dark green
to olive-buff.
a. Aerial mycelium white to ight green
to blue.
oo
246. Streptomyces — virido-
chromogenes
b. Aerial mycelium thin, white.
211. Streptomyces roseocit-
reus
ce. Aerial mycelium pale gray to blue-
gray.
49. Streptomyces chartreu-
SUS
9. Growth dark brown to black.
a. Growth on potato orange to orange-
red.
al. No aerial mycelium on potato.
200. Streptomyces purpur-
eochromogenes
b!. Aerial mycelium scant to none;
light brownish-gray.
186. Streptomyces — phaeo-
purpureus
c!. Aerial mycelium = on_ potato
abundant, gray.
10.
IE
12.
3:
198. Streptomyces purpur-
eofuscus
d'. Aerial mycelium on potato pow-
dery white.
132. Streptomyces lanatus
b. Growth on potato brown to black.
Aerial mycelium on synthetic agar
white to brownish.
at. Aerial mycelium abundant.
a®. Spirals formed.
185. Streptomyces — phaeo-
chromogenes
b?. No spirals.
153. Streptomyces melano-
genes
b!. Aerial mycelium on synthetic
agar slight.
168. Streptomyces —nobori-
loensis
Growth on synthetic media colorless
to hght orange.
a. Aerial mycelium gray to cinnamon-
drab.
26. Streptomyces aureus
b. Aerial mycelium white to gray.
192. Streptomyces poolensis
c. Aerial mycelium olive-buff.
194. Streptomyces
cundus
Growth on synthetic agar whitish-
vellow to grayish-yellow.
a. Soluble pigment light yellow.
a!. Aerial mycelium white-gray.
a®. Aerial mycelium on potato
praefe-
gray.
231. Streptomyces tana-
shiensis
b?. No aerial mycelium on po-
tato.
-oO Y oy Pee
78. Streptomyces filipinen-
Sis
bt. Aerial mycelium olive-colored.
137. Streptomyces lotdensis
b. Soluble pigment brown to reddish-
brown.
154. Streptomyces michigan-
ensis
Growth on synthetie agar gray to olive-
gray.
44. Streptomyces carnosus
Growth on synthetic media red to
purple.
a. Aerial mycelium white to gray.
71. Streptomyces erythro-
chromogenes
b. Aerial mycelium green.
14.
15.
16.
ile(s
18.
CLASSIFICATION OF STREPTOMYCES SPECIES 165
6. Streptomyces
LENSIS
afghan-
c. Aerial mycelium greenish to yellow.
4. Streptomyces acidomy-
ceticus
d. Aerial mycelium chalk-white.
59. Streptomyces collinus
Growth colorless to cream-colored.
a. Aerial mycelium cottony white,
lavender to vinaceous-lavender.
133. Streptomyces lavendu-
lae
b. Aerial mycelium
lavender.
243. Streptomyces virginiae
ec. Aerial mycelium white to cream-
colored.
grayish-pink to
172. Streptomyces odorifer
Growth yellow to brown. Aerial myce-
hum hght tan to pink.
236. Streptomyces
Growth gray to black.
79. Streptomyces
tus
venezuelae
fimbria-
Growth colorless to stone-red.
93. Streptomyces fuscus
Growth blue.
62. Streptomyces cyaneus
B. Sporophores in aerial mycelium form verticils.
I. Melanin-negative.
if.
Growth yellowish.
a. Aerial mycelium white to pinkish.
117. Streptomyces
joensis
b. Aerial mycelium greenish-yellow.
37. Streptomyces caespito-
hachi-
Sus
ce. Aerial mycelium gray.
128. Streptomyces kentuck-
ensis
Growth pink to red; aerial mycelium
pink.
76. Streptomyces fervens
Growth yellowish to green to brown;
aerial mycelium white.
149. Streptomyces mashuen-
sis
Growth colorless on synthetic media;
aerial mycelium white to light cinna-
mon.
53. Streptomyces cinnamo-
meus
Growth colorless; aerial mycelium
white.
55. Streptomyces circula-
tus
6.
Growth colorless to gray; aerial myce-
lium white to gray.
150. Streptomyces matensis
Il. Melanin-positive.
i
to
Sporophores do not produce any spi-
rals.
a. Growth white to cream-colored.
108. Streptomyces — griseo-
carneus
b. Growth colorless to yellowish.
al. Strong proteolytic properties.
a?. Aerial
mycelium on agar
media absent or white
patches.
151. Streptomyces medioci-
dicus
b?. Aerial mycelium on = agar
media white, yellowish to
gray.
84. Streptomyces flavoreti-
cult
b!. Weak proteolytic action.
a®. Aerial mycelium white to
yellowish
72. Streptomyces eurocidi-
cus
b?. Aerial mycelium
pale olive-buff.
12. Streptomyces alboniger
ce. Growth yellowish-brown.
al. Aerial mycelium white.
1. Streptomyces abikoen-
sum
b!. Aerial mycelium
yellowish tinge.
234. Streptomyces thioluteus
d. Growth dark gray to gray-green.
al. Strongly proteolytic.
238. Streptomyces verticilla-
white to
white with
tus
b!. Weakly proteolytic.
177. Streptomyces olivover-
ticillatus
e. Growth brown.
141. Streptomyces luteover-
ticillatus
Sporophores produce spirals.
a. Aerial mycelium none or limited.
232. Streptomyces tendae
b. Aerial mycelium yellow to ash-
gray.
67. Streptomyces echinatus
ce. Aerial mycelium white.
9. Streptomyces albireti-
cult
164
THE ACTINOMYCETES,
d. Aerial mycelium pale vinaceous.
161. Streptomyces netropsis
Sporophores straight or spiral-shaped.
207. Streptomyces reticuli
Verticils on secondary — branches;
growth yellowish-red to pink.
a. Spirals produced.
218. Streptomyces rubrire-
ticult
b. No spirals formed.
184. Streptomyces pentati-
CUS
Verticils on primary and secondary
branches.
a. Growth yellow to brown.
al. Aerial mycelium grayish-white.
130. Streptomyces — kitasa-
toensis
Vol.
II
b!. Growth brown to olive-drab.
176. Streptomyces — olivore-
ticult
b. Growth pink.
Cc.
120. Streptomyces hiroshi-
mensts
Growth red to reddish-brown.
214. Streptomyces roseover-
ticillatus
Sporophores may also form tufts.
a.
Aerial mycelium greenish-yellow,
turning gray.
247. Streptomyces viridofla-
vUS
. Aerial mycelium white.
229. Streptomyces spirover-
ticillatus
Cy hya-p tier
8
Description of Species of Streptomyces
the im-
portant, recognizable species of the genus
Streptomyces are given in this chapter. Most
Detailed descriptions of more
of these organisms have been isolated from
soils, composts, peats, and water basins;
some have come from dust and food ma-
terials, from plant disease lesions, and from
diseased animals and humans. Those iso-
lated from plant disease lesions may or may
not be the causative agents of such diseases;
they certainly should be considered on a par
with the soil-inhabiting forms. In the great
majority of cases, the cultures isolated from
diseased animals or from human infections
as well cannot be considered as the causative
agents of such diseases, since their patho-
genic nature has not been established ex-
perimentally.
These descriptions vary greatly both in
the details of the observations reported and
in the uniformity of treatment of such ob-
servations. For many of these observations,
the author had to depend on other com-
pilers of the literature, notably Brumpt
(1939), Lehmann and Neumann (1927),
Dodge (1935), Krassilnikov (1949), Erikson
(1935), Ettlinger et al. (1958), and others.
Unfortunately, one cannot avoid criticizing
the tendency of certain compilers to de-
scribe new species, and place others, often
well recognized and previously described
forms, as subspecies or as ‘‘also belonging
to this species,” or the even worse tendency
of some Classifiers to make certain minor
variations the basis for establishing varie-
ties of described organisms. In only
cases was an attempt made to compare newly
a few
165
isolated cultures with previously known,
although unfortunately not always avail-
able, type cultures.
Although many of the synonyms have
been examined, no detailed data are pre-
sented literature
Additional information can be found in the
latest edition of Bergey’s Manual or in the
original papers in which the descriptions
concerning references.
have appeared.
Because of the growing interest in actino-
mycetes as producers of antibiotics, numer-
ous studies of these organisms have been
made during the last 5 or 6 years. Many new
species and numerous new varieties have
been described. Old species have been better
delineated. New
have been proposed. Cooperative experi-
ments have been carried out. All this ma-
systems of classification
terial has now been critically examined, and
much additional information has been in-
cluded.
The last edition of Bergey’s Manual
(1957) contains descriptions of 150 Strepto-
The number has _ nearly
doubled in the last 5 or 6 years, as indicated
by the descriptions presented here.
myces species.
Description of Streptomyces Species
1. Streptomyces abikoensum Umezawa. et
al., 1951 TT and
Fukuyama, S. Japan Med. J. 4: 331-346,
1951; J. Antibiotics (Japan) 5: 469-476,
1952; Okami, Y. zbid. 477-480).
Morphology: Sporophores straight, short,
(Umezawa, H., Tazaki,
unbranched, bearing chains of spores. No
spirals. Certain strains produce verticils.
166
Sucrose nitrate agar: Substrate growth
yellowish-brown. Aerial mycelium — thin,
yellowish-white. Soluble pigment yellowish-
brown.
Nutrient agar: Substrate growth cream-
colored to yellow. No aerial mycelium. Solu-
ble pigment brown. Melanin-positive.
Gelatin: Growth cream-colored to brown-
ish. Soluble pigment brown. Liquefaction
crateriform.
Milk: Growth brownish. Aerial mycelium
scant, white. Soluble pigment yellowish-
brown. Peptonization, but no coagulation.
Potato: Growth wrinkled, cream-colored
to brownish. Aerial mycelium yellowish-
white. Soluble pigment reddish-brown.
Starch agar: Growth cream-colored to
yellowish. Aerial mycelium white. Hydroly-
sis good. No soluble pigment.
Nitrate reduction: Positive.
Blood agar: Growth dark cream-yellow.
Hemolysis strong.
Ege media: Growth greenish-yellow. No
aerial mycelium. Soluble pigment reddish to
violet.
Cellulose: Not decomposed.
Carbon utilization: Glucose, maltose, and
elycerol well utilized. Arabinose, xylose,
rhamnose, fructose, galactose, mannitol,
sorbitol, lactose, sucrose, raffinose, and inu-
lin not utilized.
H.S production: Negative (other strains
positive).
Tyrosinase reaction: Negative.
Antagonistic properties: Produces an
antiviral agent, abikoviromycin.
Habitat: Soil in Japan.
Remarks: Resembles S. fimicartus and
S. purpureochromogenes. Gause et al. de-
scribed a variety of this organism under the
name of A. abikoensum var. spiralis. The
above description was based upon strain
2-1-6.
* These designations represent the various cul-
ture collections where the type cultures are de-
posited. This has been elucidated in Chapter 4,
p. 78-80.
THE ACTINOMYCETES, Vol. II
Type culture: IMRU* 3654.
2. Streptomyces Nishimura
et al., 1957 (Nishimura, Hi., Kamura, I.
Tawara, K., Sasaki, K., Nakajima, K.,
Shimaoka, N., Okamoto, 8., Shimohira, M.,
and Isono, J. J. Antibiotics (Japan) 10A:
205-212, 1957).
Morphology: Sporophores long and
straight; no spirals produced. Spores oval.
Sucrose nitrate agar: Growth yellowish-
brown. Aerial mycelium well developed,
velvety, white. Soluble pigment dark yellow-
ish-brown.
Glucose-asparagine agar: Growth grayish-
olive, thin, flat; reverse pale olive. Aerial
mycelium velvety, almost white, slightly
erayish. Soluble pigment at first dull yellow,
later becoming yellowish-brown.
aburaviensis
Starch agar: Growth — grayish-yellow-
brown. Aerial mycelium grayish-white.
Soluble pigment pale yellow-brown. Hy-
drolysis weak.
Calcium malate agar: Growth pale yel-
lowish-brown. Aerial mycelium thin, white
to grayish-white. Soluble pigment grayish-
yellow-brown.
Nutrient agar: Growth thin, light gray.
No aerial mycelium. No soluble pigment.
Milk: Growth grayish-white. Aerial my-
celium white. Coagulation and peptoniza-
tion.
Potato: Growth dull yellow to pale olive.
Aerial mycelium white to light gray. No
soluble pigment.
Gelatin: Positive liquefaction. No soluble
pigment. Melanin-negative.
Nitrate reduction: Positive.
Carbon utilization: Glycerol, dextrin,
starch, glucose, maltose, galactose, inulin,
and fructose utilized. Mannitol, arabinose,
inositol, xylose, and
rafinose, a-lactose,
sucrose not utilized.
3. Streptomyces achromogenes Okami and
Umezawa, 1953 (Umezawa, H., Takeuchi,
T., Okami, Y., and Tazaki, T. Japan. J:
Med. Sci. Biol. 6: 261-268, 1953).
DESCRIPTION OF SPECIES OF STREPTOMYCES
Morphology: Sporophores straight, no
spirals. Spores cylindrical.
Glycerol nitrate agar: Growth colorless to
brownish. Aerial mycelium scant, white to
dark grayish. Soluble pigment brown.
Glucose-asparagine agar: Growth yellow-
ish brown. Aerial mycelium scant, yellowish-
white. Soluble pigment none or slightly
brown.
Nutrient agar: Growth wrinkled, elevated,
colorless to brownish. No aerial mycelium
or soluble pigment.
Potato: Growth — yellowish-brown — to
brownish, fine, wrinkled. Aerial mycelium
white, powdery. Soluble pigment absent at
first, later reddish-brown.
Gelatin: Growth yellowish-brown. Soluble
pigment shghtly brown. Liquefaction very
weak. Melanin-negative.
Milk: Surface growth poor. No soluble pig-
ment. Coagulation and slow peptonization.
Egg media: Growth — reddish-brown,
wrinkled. No aerial mycelium. No soluble
pigment.
Nitrate reduction: Positive.
Antagonistic properties: Produces an anti-
viral agent, achromoviromycin.
Remarks: This culture resembles S.
diastaticus and S. fimicarius. It is character-
ized by the brown pigmentation on syn-
thetic agar only. A strain of this organism,
which produces the antibiotic streptozotocin,
was isolated by Vavra et al. (1959) from a
soil in Kansas; they have further cultural
data concerning the original culture and the
new strain.
Type culture: IMRU 3730; ATCC 12,767.
4. Streptomyces acidomyceticus Ogata et al.,
1954 (Ogata, K., Miyake, A., and Morimoto,
A. Japanese Patent No. 204,403, March 5,
1954).
Morphology: Sporophores usually do not
form spirals. Spores cylindrical or oval,
0.8 to 1.2 by 1.4 to 1.8 u.
Sucrose first
ereenish-brown.
nitrate agar: Growth at
light yellow, later dark
167
Aerial mycelium greenish to yellow-white.
Soluble pigment — slightly
sometimes absent.
violet-colored:
Glucose-asparagine agar: Growth brown-
ish-yellow or brownish-red, and_ partially
greenish-blue.
Calcium malate agar: Aerial mycelium
Soluble
sometimes absent.
Glucose nutrient agar: Growth brownish,
partially dark blue. Aerial hyphae gray-
white. Soluble pigment brownish-black.
Gelatin: Growth dark green. Aerial my-
celium greenish-white. Soluble pigment
greenish-brown. Liquefaction limited.
Potato: Growth greenish-brown. Aerial
mycelium at first white, later pinkish-red.
Soluble pigment dark green.
Milk: Growth cream-colored, later turning
light brown. No coagulation. Soluble pig-
ment light brown.
Starch: Slow decomposition.
Tyrosinase reaction: Negative.
Nitrate reduction: Positive.
Production of H.S: Positive.
Carbon utilization: Arabinose, glucose,
maltose, lactose, salicin, and salts of or-
ganic acids utilized. Xylose, fructose, raffi-
nose, inulin not attacked.
Antagonistic properties:
antibiotic acidomycin.
Remarks: S. acidomyceticus is closely re-
lated to S. phaeochromogenes, the latter
ereenish-white. pigment violet;
Produces the
forming spirals in gelatin media, but not the
former.
Type culture: ATCC 11,611.
5. Streptomyces acidophilus (Jensen, 1928)
Waksman and Henrici, 1948 (Jensen, H. L.
Soil Sci. 25: 226, 1928).
Morphology: Sporophores either few or
numerous, with sinistrorse spirals. Spores
oval and spherical, 1.0 to 1.2 by 1.2 to 1.5
LL.
Agar media: Growth on acid media (pH
2.0 to 6.0) colorless. Aerial mycelium whitish.
Sucrose nitrate agar: No growth.
168
Glucose-asparagine agar: Growth raised,
somewhat wrinkled, young
cultures. Aerial mycelium thin, white at
colorless in
first, later gray or yellowish-brown.
Nutrient agar: No growth.
Starch agar: Growth at 25°C good, color-
less. Aerial mycelium abundant, smooth,
white. Some diastatic action.
Potato: Growth good, raised, folded. No
discoloration of plug. Melanin-negative.
Gelatin: Growth after 10 days very scant,
thin, semitransparent, colorless. Liquefaction
slow.
Milk: No growth.
Nitrate reduction: Trace.
Sucrose: No inversion.
Antagonistic properties: Strongly positive.
Habitat: Soil.
Remarks: Grows in acid media only, with
an optimum at pH 3.5 to 4.5.
6. Streptomyces afghaniensis Shimo et al.
1959 (Shimo, M., Shiga, T., Tomosugi, T.,
and Kamoi, I. J. Antibiotics (Japan) 12A: 1,
1959).
Morphology: Sporophores form spirals.
nitrate agar: Growth — olive-
colored, with reddish-brown reverse. Aerial
mycelium pale green to light greenish-gray.
Soluble pigment brown to reddish-brown.
Sucrose
Glucose-asparagine agar: Growth olive-
colored, with reddish-brown reverse. Aerial
mycelium pale green to light greenish-gray.
Soluble pigment brown to reddish-brown.
Calcium malate agar: Growth olive-
colored. Aerial mycelium pale yellow-orange
to pale orange. Soluble pigment yellowish-
brown to reddish-brown.
Nutrient agar: Growth colorless to olive
to buff. Aerial mycelium grayish-white.
Soluble pigment light brown.
Gelatin: Growth colorless. Aerial my-
celium white. Soluble pigment brown.
Medium liquefaction.
Milk: Growth yellowish-brown. No aerial
mycelium. Soluble pigment brown to dark
brown.
THE ACTINOMYCETES, Vol. II
Potato: Growth wrinkled, colorless. Aerial
mycehum olive to yellowish-brown. Soluble
pigment yellowish-brown.
Cellulose: Positive growth.
Nitrate reduction: Negative.
Tyrosinase: Doubtful.
Carbon source: Utilizes rhamnose, raffi-
nose, and other carbohydrates; does not
utilize sodium citrate and sodium acetate;
doubtful growth on dulcitol and sorbitol.
Antagonistic properties: Produces an
antibiotic, taitomycin, active upon gram-
positive bacteria.
Habitat: Soil in Afghanistan.
Remarks: Resembles S. collinus and S.
erythrochromogenes.
7. Streptomyces albidoflavus (Rossi-Doria,
1891, emend. Gasperini, 1894) Waksman and
Henrici, 1948 (Rossi-Doria, T. Ann. ist. ig.
sper. Roma, n. s. 1: 399-438, 1894).
Synonym: Actinomyces albido-flavus
Duché, 1934, emend. Krassilnikov, 1949.
Morphology: Sporophores short, spiral-
forming, sinistrorse. Spores spherical.
Glucose-asparagine agar: Growth brown.
Aerial mycelium white, later becoming
whitish-vellow. Soluble pigment yellowish.
agar: Growth cream-
colored, covered with fine aerial
mycelium; yellow soluble pigment.
Tyrosine agar: Growth fine with orange-
yellow on reverse side; medium becomes
Glucose-peptone
white
vellowish to yellowish-rose.
Gelatin: Punctiform colonies with white
aerial mycelium on surface. No soluble
pigment. Rapid liquefaction.
Milk: Growth rapid, becoming covered
with whitish aerial mycelium; never fully
covering the surface; no coagulation; pep-
tonization begins slowly and is completed in
13 days; liquid colored yellowish-orange.
Starch media: Growth
covered with yellow aerial mycelium. After
20. days, growth becomes much _ folded;
greenish on reverse side; soluble pigment
cream-colored,
shghtly amber. Hydrolysis.
DESCRIPTION OF
Cellulose: Some growth.
Coagulated serum: Cream-colored growth
on surface. Aerial mycelium white. Liquefac-
tion rapid.
Production of HS: Negative.
Antagonistic properties: Produces strep-
tothricin.
Habitat: Soil.
Remarks: According to Flaig and Kutz-
ner (1960), this culture obtained from CBS
is S. coelicolor Miller. Ettlinger et al. (1958)
considered that Duché’s strain of this
organism belongs to S. griseus.
8. Streptomyces albidus (Duché, 1934)
Waksman (Duché, J.
groupe albus. P. Lechevalier, Paris, 1934).
form
Les actinomyces du
Morphology: Sporophores long,
open spirals. Spores spherical to oval.
Glucose nitrate agar: Growth colorless;
some drops of colorless guttation. Aerial
mycelium white. Soluble pigment yellowish.
Peptone agar: Growth colorless. Aerial
mycelium white; reverse slightly greenish.
Soluble pigment brownish.
Potato: Growth flat, colorless. Aerial
mycelium white. No soluble pigment.
Gelatin: Growth cream-colored. Rapid
liquefaction. No soluble pigment. Melanin-
negative.
Milk: Growth cream-colored. Coagulation
weak; peptonization rapid. Odor cheesy.
Starch: Hydrolysis good.
Cellulose: Growth good.
Fats and waxes: Growth good, according
to Krassilnikov (1949).
Nitrate: Slow reduction to nitrate.
Odor: Strong, earthy.
properties:
Krassilnikov (1949), it
antagonistic activities.
Remarks: Closely related to S.
(Krassilnikov, 1949) ; differs by more delicate
growth, by a reverse that is often yellowish-
brown. Also related to S. microflavus, but
differs from the form described by Krainsky
in that its growth is never rose-yellow and
Antagonistic According to
possesses strong
albus
SPECIES OF STREPTOMYCES
169
that it grows abundantly on potato. Gause
et al. (1957) described a variety of this
organism under the name A. albidus var.
invertens. Exttlinger et al. (1958) considered
it as a strain of S. griseus.
9. Streptomyces albireticuli Nakazawa,
1955 (Nakazawa, K. J. Agr. Chem. Soc.
Japan 29: 644-647; 647-649, 1955).
Morphology : the
secondary verticils of the aerial mycelium.
The spores are cylindrical 0.6 to 0.8 by 1.4
LO eS a:
Sucrose nitrate agar: Growth thin, color-
Produces spirals in
less; reverse pale ochraceous salmon. Aerial
mycelium white.
Glucose-asparagine agar: Growth colorless;
becoming yellow. Aerial mycelium
white, cottony, later becoming cream-colored.
later
Nutrient agar: Growth thin, mouse-gray.
No aerial mycelium. Soluble pigment och-
raceous tawny.
Potato plug: Growth gray. Aerial my-
celium white. Color of plug brown.
Gelatin: Liquefaction slow. Soluble pig-
ment black.
Milk: Growth cream-colored. Peptoniza-
tion slow. Soluble pigment brown after 24
days.
Starch: Actively diastatic.
Nitrate reduction: Positive.
Production of H.S: Positive.
Cellulose: No growth.
Antagonistic properties: Produces euro-
cidin, an antifungal antibiotic.
10. Streptomyces alboflavus (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, 8. A. and Curtis, R. E. Soil Sci.
1: 99-134, 1916; 8: 90, 1919).
Morphology: Sporophores straight, branch-
ing, with very little tendency to produce
spirals. Spores oval-shaped.
Sucrose nitrate Growth — glossy,
agar:
spreading, colorless, becoming yellowish.
Aerial
yellowish tinge. No soluble pigment.
mycelium powdery, white, with
170
Glycerol malate agar: Growth light
pinkish-cinnamon. Aerial mycelium late,
white.
Glucose-asparagine agar: Growth — re-
stricted, much folded, cream-colored with
sulfur-yellow surface. No aerial mycelium.
No soluble pigment.
Nutrient agar: Growth restricted, cream-
colored. No aerial mycelium. No soluble
pigment.
Potato: Growth wrinkled, moist, cream-
colored.
Gelatin: Surface growth abundant, color-
less. Aerial mycelium white or absent. No
soluble pigment. Slow liquefaction.
Milk: Surface ring pinkish. No coagula-
tion; limited peptonization.
Starch media: Growth thin, spreading,
yellowish. No aerial mycelium. Good hy-
drolysis of starch.
Cellulose: Scant growth.
Nitrate reduction: Positive.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Positive.
Remarks: Various cultures related to this
organism have been described under a
variety of different names. It is sufficient to
mention A. cremeus, A. griseoloalbus, A.
flavidovirens, and a variety of the latter,
fuscus, described by Gause et al. (1957).
KrassiInikov (1949) considered it as a
variety of A. flavus.
Type culture: IMRU 3008.
11. Streptomyces albogriseolus Benedict. et
al., 1954 (Benedict, R. G., Shotwell, O. L.,
Pridham, T. G., Lindenfelser, L. A., and
Haynes, W. C. Antibiotics & Chemotherapy
4: 653-656, 1954).
Morphology: Sporophores monopodially
branched, producing short, compact spirals,
averaging 4 to 6 turns. Spores spherical or
oval, covered with numerous long, fine hairs
(PL ln):
Sucrose Aerial mycelium
nitrate agar:
THE ACTINOMYCETES, Vol. II
white, becoming ash-gray, often with white
spots.
Starch agar: Aerial mycelium white to
dark gray. Hydrolysis.
Nutrient agar: Aerial mycelium white to
ash-gray. Melanin-negative.
Potato: Growth cretaceous to dirty gray-
ish-white to faint pink.
Carrot: Vegetative growth white to dirty
cream; no aerial mycelium. Slant not dark-
ened.
Gelatin: Dirty white sediment. Positive
liquefaction. Not pigmented.
Milk: Orange-colored ring; partially pep-
tonized at 14 days.
Nitrate: Reduction to nitrite.
Production of H.S: Negative.
Temperature: Good growth at 25-41°C.
No growth at 50°C.
Antagonistic properties: Produces a ‘“‘neo-
mycin complex.”
Habitat: Soil.
Type culture: IMRU 3698.
12. Streptomyces alboniger Hesseltine et al.,
1954 (Hesseltine, C. W., Porter, J. N.,
Deduck, N., Hauck, M., Bohonos, N., and
Williams, J. H. Mycologia 46: 16-23, 1954).
Morphology: Sporophores — irregularly
branched, erect to flexuous; no spirals. Ver-
ticils produced. Spores catenulate, oval,
0.8 by 1.25 yp.
Sucrose nitrate agar: Growth poor, white.
Aerial mycelium white to pale olive-buft.
No soluble pigment.
Glucose-asparagine agar: Growth black-
ish-gray. Aerial mycelium white. Soluble
pigment blackish-gray.
Nutrient agar: Growth moist, smooth,
colorless to
black. No
pigment.
Starch agar: Growth good. Aerial my-
celium white to pale olive-buff. Soluble
yellowish, to dark brown or
aerial mycelium. No soluble
pigment black. Good hydrolysis.
Potato: Growth moist, yellow. Aerial
DESCRIPTION OF SPECIES OF STREPTOMYCES 171
mycelium white. Soluble pigment dark,
greenish-black.
Gelatin: Growth fair. Aerial mycelium
white. Soluble pigment light yellow. Lique-
faction medium.
Milk: Surface white ring, with yellow-
green to light yellow-brown below surface.
Aerial mycelium white. Peptonization slow.
Cellulose: No growth.
Production of HoS: Negative.
Antagonistic properties: Produces puro-
mycin, an antibiotic active upon certain
gram-positive bacteria and protozoa.
Habitat: Forest soil.
Remarks: Culture is characterized by the
formation of an olivaceous black
pigment in some media, such as asparagine-
soluble
glucose agar, but no such pigment is pro-
duced on certain organic media.
Type culture: ATCC 12,461.
13. Streptomyces albosporeus (Krainsky,
1914) Waksman and Henrici, 1948 (Krain-
sky, A. Centr. Bakteriol. Parasitenk. Abt.
II., 41: 687, 1914: Waksman, S. A. and
Curtis, R. E. Soil Sci. 1: 99, 1916; 8: 90,
1919).
Morphology : straight,
branching, with occasional spirals. Spores
spherical or oval, 0.8 to 1.2 by 1.0 to L.8u.
Sucrose nitrate agar: Growth spreading,
Sporophores
colorless, with pink center, becoming brown-
ish, Aerial mycelium white,
covering the whole surface; often none. No
vinaceous.
soluble pigment.
Glycerol malate agar: Growth rose to
orange-red. Aerial mycelium white, later
changing to yellow. No soluble pigment.
Glucose-asparagine agar: Growth wrinkled,
spreading, red, with colorless margin. Aerial
mycelium appears late, white.
Nutrient agar: Small, cream-colored col-
omies. No aerial mycelium. No soluble
pigment.
Starch agar: Growth thin, spreading,
transparent, with red tinge. No aerial
mycelium. Ready hydrolysis.
Potato: Growth red to brownish-gray.
No aerial mycelium, or Melanin-
negative.
white.
Gelatin: Growth yellow, changing to red,
with hyaline margin. Usually no aerial my-
celium; when produced, sometimes gray.
Medium liquefaction.
Milk: Seant, pink ring. No coagulation;
no peptonization.
Cellulose: No growth or scant.
Nitrate reduction: Fair.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Positive.
Habitat: Soil.
Remarks: Above description is based
partly upon the isolates of Waksman and
Curtis, since Krainsky’s culture was not
available. Krassilnikov (1949) considered it
as a variety of A. ruber. According to
Ettlinger et al. (1958) this organism should
be considered as a strain of S. griseus, a
hardly justifiable assumption.
Type culture: IMRU 3003.
14. Streptomyces alboviridis (Duché, 1934)
Waksman (Duché, J. Les actinomyces du
groupe albus. P. Lechevalier, Paris, p. 317,
1934).
Morphology: According to Krassilnikov
(1949), the sporophores produce spirals with
3 to + turns. Spores spherical.
nitrate Growth
Glucose agar:
colored becoming olive-green. Aerial myce-
cream-
lum white, becoming yvellowish-green. Solu-
ble pigment brownish.
Glucose-asparagine agar: Growth at first
white, becoming olive-colored to almost
dark. Aerial mycelium white to green. Solu-
ble pigment yellowish.
Starch agar: Growth cream-colored; re-
verse brownish-green. Aerial mycelium
white, becoming green.
Gelatin: Growth white, becoming green.
Soluble pigment greenish-brown. Liquefac-
tion rapid (slow, according to Krassilnikov,
1949).
172 THE ACTINOMYCETES, Vol. II
Potato: Growth white, becoming brown-
ish to rust-colored. Plug colored black.
Tyrosine agar: Growth white with a
brownish reverse. Soluble pigment brownish.
Milk: Coagulation and peptonization.
Coagulated serum: Growth cream-colored.
No aerial mycelium. No soluble pigment.
Rapid liquefaction of serum.
Remarks: This organism is considered as
a transitional form between S. albus and S.
viridis. Krassilnikov (1949) considered it as
a variety of A. viridochromogenes.
15. Streptomyces albus (Rossi-Doria, 1891;
emend. Gasperini, 1892) Waksman and Hen-
rici, 1948 (Rossi-Doria, T. Ann. ist. ig. sper.
Roma, n. s. 1: 399-438, 1894).
Synonym: Numerous synonyms of this
species are found in the literature. They
belong mostly to the species-group ““S. albus.”
Many of them are listed in Chapter 6, under
the corresponding group.
Morphology: Sporophores produce long
spirals. Spores spherical to oval. Some strains
produce, according to Okami, straight sporo-
phores, depending on the composition of the
medium.
Agar media: Growth colorless; may be-
come yellowish to brown with age. No
pigment formed, although
strains may excrete a brownish substance in
certain media and under certain conditions.
Aerial mycelium abundant, white; the shade
of color varies with composition of medium
soluble some
from snow-white to somewhat yellowish.
Sucrose nitrate agar: Substrate growth
smooth, colorless. Aerial mycelium cottony
to powdery; white to snow-white.
Glucose-asparagine agar: Aerial mycelium
gray, becoming brownish.
Nutrient agar: Generally no aerial my-
celium; chalky white deposit on old colonies.
Potato: Growth lichenoid, cream-colored.
Aerial mycelium white.
Gelatin: Colonies gray. No soluble pig-
ment. Strong liquefaction.
Milk: Surface ring cream-colored. Aerial
mycelium white. Peptonization rapid.
Starch agar: Aerial mycelium white.
Rapid hydrolysis of starch in some cultures;
others show little or no hydrolysis.
Nitrate: Reduction to nitrite positive.
Production of H.S: Negative.
Odor: Characteristic, moldy.
Antagonistic properties: Certain strains
are active upon gram-positive bacteria. Some
produce actinomycetin, others form thiolutin
or endomycin.
Habitat: Occurs in dust and soil.
Xemarks: The general occurrence of this
species, the ease of its superficial identifica-
tion, and the fact that it has been adopted
as the type species for the genus Strepto-
myces, Justify a more complete characteri-
zation, as given in Chapter 6. Numerous
strains of this species, varying in. their
cultural and other properties have been
reported. Numerous descriptions of closely
related organisms also are found in the
literature (Duché). IXrassilnikov lists 18
strains and substrains (4. albus vulgaris, A.
albus chlamydosporus, etc.). A. longisporus
KrassilInikov (1949) and some of the sub-
strains, like A. longisporus griseus, belong to
this group. Solovieva and Rudaya (Anti-
biotiki, 4(6): 5-10, 1959) list a variety fun-
gatus capable of producing an antifungal
agent, albofungin.
Type culture: IMRU 3005.
16. Streptomyces althioticus Yamaguchi
et al., 1957 (Yamaguchi, H., Nakayama,
Y., Takeda, K., Tawara, K., Maeda, K.,
Takeuchi, T., and Umezawa, H. J. Anti-
biotics (Japan) LOA: 195-200, 1957).
Morphology: Curved chains or spirals of
oval spores on ends of aerial sporophores.
Frequently, tips of aerial mycelium divided
into tufts of spore chains.
Sucrose nitrate agar: Growth colorless to
white, later light brown to purplish. Aerial
mycelium powdery white, later gray.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Glucose-asparagine agar: Growth color-
less to white, later light brown with or
without dull light reddish tinge. Surface
glossy. Aerial mycelium scant, white. Light
brown to dull light reddish-brown soluble
pigment.
Starch agar: Aerial mycelium white to
eray. No soluble pigment. No hydrolysis in
7 days.
Glucose-asparagine agar: Growth color-
less to white, later light brown with or with-
out dull light reddish tinge. Surface glossy.
Aerial mycelium scant, white. Light brown
to dull light reddish-brown soluble pigment.
Glucose nutrient agar: Growth heht
yvellowish-brown. Surface glossy. Aerial
mycelium white to gray. Soluble pigment
light yellowish-brown.
Potato: Growth abundant, light yellowish-
brown. Aerial mycelium white to gray.
Gelatin: Scant growth. No liquefaction.
Brown soluble pigment.
Milk: Light yellowish-brown surface ring,
with scant white aerial mycelium. Yellow-
ish-orange soluble pigment occasionally.
Peptonization positive.
Egg medium (37°C): Growth yellow with
eray tinge. Aerial mycelium white, later
light purplish occasionally.
Cellulose: Scant growth with purplish-
gray aerial mycelium and light purplish
pigment.
Carbon utilization: Abundant growth with
rhamnose, fructose, galactose, mannitol, and
glucose; weak growth with xylose, arabinose,
maltose, sorbitol, and inositol; none = or
very scant with dulcitol, raffinose, and
inulin.
Antagonistic properties: Produces anti-
biotic althiomycin.
Remarks: Closely related to S. achromo-
genes and S. rimosus. Spiral formation of
culture, no nitrite formation, and purplish
tone of growth and aerial mycelium differ-
entiate it from S. achromogenes. Purplish
tinge of aerial mycelium and growth, soluble
Lis
pigment, no nitrite formation, and no
cracked surface of the growth differentiate
it from S. rimosus.
17. Streptomyces ambofaciens Pinnert-Sin-
dico, 1954 (Pinnert-Sindico, S. Ann.
Pasteur 87: 703-707, 1954).
Morphology: Sporophores form spirals.
Spores oval or spherical.
inst.
Sucrose nitrate agar: Substrate growth
yellow to gray. Aerial mycelium white to
gray. Soluble pigment weak brownish-yel-
low.
Glucose-asparagine agar: Growth yellow,
covered with white aerial mycelium. Soluble
pigment weak yellow-brown.
Calcium malate agar: Growth similar to
that on sucrose nitrate agar. No soluble
pigment.
Potato: Growth clear brown. Aerial
mycelium powdery gray. Soluble pigment
weakly brown to brownish-red.
Gelatin: Surface growth yellow; flakes in
liquefied portion. Medium
Weak brown-orange pigment in liquefied
zone. Melanin-negative.
Milk: No coagulation, partial peptoniza-
tion in | month. Peptonized zone orange-
liquefaction.
brown to red.
Nitrate: Weak
synthetic media; none at all in organic
media.
Production of HoS: Negative.
Carbon utilization: Glycerol, arabinose,
reduction to nitrite in
glucose, galactose, levulose, mannose, lac-
tose, rhamnose, starch, and mannitol well
utilized. Raffinose, erythritol, dulcitol, and
sorbitol not utilized.
Antagonistic properties: Produces two
antibiotics, congocidin and spiramycin.
Remarks: Ettlinger ef al. (1958) included
this organism with S. auwreofaciens.
(Beijerinck,
1912; emend. Krassilnikov, 1941) Waksman
18. Streptomyces annulatus
(Krassilnikov, N. A. Actinomycetales. Izvest.
Akad. Nauk, SSSR, Moskau, 1941).
174
Not A. annulatus Wollenweber, 1920.
Morphology: Sporophores produce spirals,
with 3 to 7 turns (sinistrorse). Spores spheri-
=
cal, 0:7 bm.
Sucrose nitrate agar: Growth colorless,
flat, penetrating deep into agar. Aerial
mycelium white, velvety, growing in the
form of concentric rings.
Nutrient agar: Colorless growth. Aerial
mycelium white, concentric rings — less
marked. Melanin-negative.
Gelatin: Slow hquefaction.
Milk: Positive coagulation and slow
peptonization.
Starch: Hydrolysis.
Cellulose: Growth good.
Invertase: Positive.
Production of HS: Negative.
Odor: Strong, earthy.
Antagonistic properties: Highly antag-
onistic to mycobacteria and gram-positive
bacteria; some strains are active against
fungl.
Habitat: Soil.
Remarks: Krassilnikov (1949)
this organism as a variety of S. albus.
Type culture: IMRU 3307.
19. Streptomyces antibioticus (Waksman
and Woodruff, 1941) Waksman and Henrici,
1948 (Waksman, 8S. A. and Woodruff, H. B.
J. Bacteriol. 42: 232, 246, 1941; see also
Waksman, 8. A. and Gregory, F. J. Anti-
biotics & Chemotherapy 4: 1050-1056, 1954).
Morphology: Sporophores straight, long,
considers
arranged in clusters or broom-shaped bodies;
usually not wavy and no spirals; some
strains may produce a few spirals. Spores
nearly spherical to somewhat elliptical,
smooth (Pl. II 1). Capacity to produce aerial
mycelium lost upon continued
cultivation on artificial media (Pl. V Fb).
Sucrose nitrate
may be
agar: Growth cream-
colored to yellowish, tending to darken in
reverse. Aerial mycelium light to mouse-
gray, with white patches. Soluble pigment
faint yellowish to yellow to dark.
THE ACTINOMYCETES, Vol. II
Glucose-asparagine agar: Growth cream-
colored, with yellowish to orange to dark
reverse. Aerial mycelium light to ash-gray.
Soluble pigment absent or yellow to brown-
ish.
Calcium malate agar: Growth colorless to
yellowish. Aerial mycelium white to white-
eray.
Nutrient agar: Growth brownish, thin.
Aerial mycelium yellowish-gray to yellow-
ish-green. Soluble pigment brown to dark.
Melanin-positive.
Potato: Growth thin to heavy, lichenoid;
brownish to orange in color, sometimes
olive-green. Aerial mycelium absent or
thin to patchy, white or gray. Soluble pig-
ment brownish to dark; absent in many
cultures.
Gelatin: Growth yellowish to brown to
dark brown. Aerial mycelium as patches of
white to gray. Soluble pigment black.
Liquefaction at first very slow, later be-
coming more rapid.
Milk: Thick surface ring, brownish. Aerial
mycelium mouse-gray with greenish tinge.
No coagulation, but gradual peptonization.
Soluble pigment brownish to black.
Production of HS: Positive.
Tyrosinase: Negative.
Antagonistic properties: Marked antag-
onistic effect on bacteria and fungi. Produces
actinomycin A, the first crystalline anti-
biotic ever isolated from an actinomycete
culture.
Source: Isolated from soil on Escherichia
coli-washed plate, using living cells of F.
coli as the only source of available nutrients.
Later also isolated from a variety of dif-
ferent soils.
Remarks: Ettlinger et al. (1958) included
in this group S. bikiniensis, S. cinereoruber,
S. eurythermus, and S. tpomoeae. Krassil-
nikovy (1949) included this species with A.
parvus.
Type culture: IMRU 3435.
20. Streptomyces antimycoticus Waksman
DESCRIPTION OF SPECIES OF STREPTOMYCES 175
(Leben, C., Stessel, G. J., Keitt, G. W.
Mycologia 44: 159-169, 1952).
Morphology:
situated typically in dense groups. Spirals
tend to be open, becoming closed and com-
pact prior to the formation of spores. Spores
oval, 0.6 to 1.3 by 0.7 to 2.0 yu.
Sucrose
at first white, later gray. Aerial mycelium
abundant, light neutral gray. No soluble
Sporophores with spirals
nitrate agar: Substrate growth
pigment.
Glycerol malate agar: Aerial mycelium
abundant, light neutral gray. Soluble pig-
ment faint green.
Nutrient peptone agar: Growth shiny,
cream-colored. Aerial mycelium moderate,
pebbly, white. No soluble pigment. Melanin-
negative.
Potato-glucose agar: Aerial
abundant, neutral gray. Soluble pigment
mycelium
faint, brown.
Yeast extract Aerial
abundant, neutral gray. No soluble pigment.
Starch agar: Aerial mycelium abundant,
white to neutral gray. No soluble pigment.
Diastatic action weak to moderate.
Potato: Growth finely wrinkled, cream-
colored. Aerial mycelium sparse. Plug dark-
ened slightly.
Gelatin: Growth — translucent,
colored. Aerial mycelium sparse,
Liquefaction slight at 15 days, moderate
at 30 days. No soluble pigment.
Milk: Ring cream-colored. Coagulation;
peptonization in 15 to 30 days. Yellowish-
agar: mycelium
cream-
white.
orange pigmentation.
Nitrate reduction: Shght.
Antagonistic properties: Produces an anti-
fungal agent, helixin.
21. Streptomyces arenae Grundy, 1954
(Grundy, W. E. Brit. Pat. 719,230, Dec. 1,
1954*).
Morphology: Monopodial branching of
mycelium. Sporophores terminate in tight
spirals. Spores spherical to oval.
* Supplemented by personal communication.
Sucrose nitrate agar: Growth wrinkled,
vellow, turning dark orange-brown with age.
Aerial mycelium grayish-white. Soluble pig-
ment light yellow-brown.
Calcium malate
colored, turning bright reddish-brown with
agar: Growth cream-
age. Aerial mycelium fluffy, cream-colored
turning gray with pink tinge. Soluble pig-
ment light pink. Complete dissolution of the
calcium malate.
Glucose-asparagine agar: Growth sparse,
golden brown; a few tufts of white aerial
mycelium. Soluble pigment yellow.
Nutrient agar: Growth moderate, golden
brown. Aerial mycelium gray-white, spores
turning darker gray. Soluble pigment light
brown.
Potato: Growth abundant, golden brown,
turning dark brown. Aerial mycelium fluffy,
becoming on sporulation dark gray. Potato
dark gray, turning black.
Gelatin: Heavy gray pellicle on surface.
Liquefaction slow. Soluble pigment deep red-
brown diffusing through the liquefied zone.
Medium liquefaction.
Milk: Heavy pellicle. Milk digested in 25
to 28 days with formation of curd just before
complete digestion. Soluble pigment dark
brown, turning black in 30 days.
Starch: Hydrolysis slow.
Nitrate: No reduction.
Carbon utilization: Good growth with
xylose, glucose, mannose, galactose, lactose,
maltose, sucrose, starch, mannitol, glycerol
sodium acetate, sodium citrate, and potas-
sium sodium tartrate. Sorbitol and calcium
lactate not utilized.
Antagonistic properties: Produces an anti-
biotic active upon Mycobacterium tuberculo-
Sts.
Habitat: Illinois soil.
22. Streptomyces argenteolus Perlman,
1957 (Perlman, D. U. S. Patent 2,709,705,
October 7, 1958).
Morphology: Aerial
generally branched, not forming loops or
mycelium hyaline,
176
spirals; individual filaments are rarely sep-
tate. straight, flexuous, or
fascicled (in tufts). Spores are oval to oblong,
1.0 to 1.2 uw. The spore color is ight gull-gray.
Sucrose nitrate agar: No growth.
Nutrient agar: Growth colorless, abun-
Sporophores
dant, spreading. Aerial mycelium white. No
soluble pigment.
Oatmeal agar: Growth good. Aerial myce-
lium limited, no sporulation. Soluble pig-
ment shght maize-yellow.
Casein digest-meat extract agar: Growth
abundant, dark olive-buff. Aerial mycelium
well developed, pale smoke-gray. No soluble
pigment.
Gelatin: Rapid liquefaction. Melanin-neg-
ative.
Milk: Positive coagulation and peptoniza-
tion.
Potato: Growth good, creamy-buff, cere-
briform. Aerial mycelium white; no sporula-
tion. No soluble pigment.
Starch: Hydrolyzed.
Nitrate reduction: Positive.
Carbon utilization: Mannitol, d-xylose,
l-arabinose, /-rhamnose, d-fructose, treha-
lose, and lactose utilized. No growth or very
scant growth with inositol, sorbitol, meli-
biose, sucrose, and dextrin.
Habitat: Soil.
Biochemical activities: Certain strains of
this organism convert progesterone to 16
a-hydroxyprogesterone.
23. Streptomyces armillatus Mancy-Cour-
tillet and Pinnert-Sindico, 1954 (Mancy-
Courtillet, D. and Pinnert-Sindico, S. Ann.
inst. Pasteur 87: 580-584, 1954).
Morphology: Aerial mycelium produces
spirals.
Glucose- or glycerol-asparagine
Growth yellow-gray. Aerial mycelium poorly
agar:
developed, white.
Sucrose nitrate agar: Growth very poor,
colorless. No aerial mycelium.
Glucose nitrate agar: Growth poor. No
soluble pigment. No reduction of nitrate.
THE ACTINOMYCETES, Vol. II
Glucose-peptone agar: Growth very good,
vellow-gray. Aerial mycelium poorly devel-
oped, white. Soluble pigment weak rose,
becoming brownish. Melanin-negative.
Potato: Growth good, yellow-gray. Aerial
mycelium
pigment.
poorly developed. No soluble
Tyrosine medium: Growth flat, yellow-
gray, becoming beige. Aerial
white. No soluble pigment.
Gelatin: Growth in form of pellicle. Aerial]
mycelium white. Soluble
brown. Rapid liquefaction.
Milk: Growth in form of surface ring,
mycelium
pigment rose-
eray to yellow. Peptonized portion colored
yellow. Coagulation and rapid peptoniza-
tion.
Starch: No hydrolysis.
Antagonistic properties: Produces oxytet-
racyclhine.
temarks: This organism can be classified
with the S. bobiliae-S. erythreus group, al-
though its growth is yellow rather than red.
It grows poorly upon synthetic media, upon
which it forms no aerial mycelium. It does
not reduce nitrate to nitrite. It does not
produce a purple pigment upon egg media.
It does not change the reaction of milk to
alkaline. It does not hydrolyze starch. It
differs from S. rimosus and S. griseoflavus,
which produce yellow to brown pigments;
S. armillatus under the same conditions does
not form any pigment.
24. Streptomyces aurantiacus — (Rossi-
Doria, 1891 emend. Gasperini, 1892; emend.
Waksman
A. Actinomycetales. Izvest. Akad.
SSSR, Moskau, p. 36, 1941).
Morphology: Produces an abundance of
(Krassilnikov, N.
Nauk.
Kkrassilnikov )
chlamydospores. Sporophores form. spirals
with 3 to 5 turns. Spores spherical to oval,
0.7 to 0.9 by 0.6 to 0.8 u.
Agar media: Growth lichenoid, dry, com-
pact; colored bright orange or golden. Pig-
ment insoluble in medium, but soluble in
DESCRIPTION OF SPECIES OF STREPTOMYCES 177
organic solvents. Aerial mycelium poorly
developed. Melanin-negative.
Potato: Soluble pigment brown.
Gelatin: Growth yellow to orange-yellow
to deep orange. Liquefaction none or slow.
No aerial mycelium.
Milk: Surface growth orange. No coagula-
tion; unchanged or weak peptonization.
Starch: Slow hydrolysis.
Cellulose: No growth.
Nitrate: No reduction.
Invertase: None.
Fats: Hydrolysis and utilization rapid.
Paraffin: Growth good, with spiral-form-
ing sporophores and spherical spores.
Pigment: Red-orange, extracted with 96
per cent alcohol. The orange pigment was
dissolved in petroleum ether, the red pig-
ment being insoluble (Ixriss).
Antagonistic properties: Strongly antago-
nistic to gram-positive bacteria.
Habitat: Soil, dust.
Remarks: Some strains deposit ferric
oxide on the surface of the hyphae.
25. Streptomyces aureofaciens Duggar,
1949 (Duggar, B. M. Ann. N. Y. Acad. Sci.
51: 177, 1948; U.S. Patent 2,482,055, Sept.
14, 1949).
Morphology: Sporophores monopodially
branched, flexuous, producing open spirals.
Spores spherical to oval, smooth (PI. II m).
Sucrose nitrate agar: Substrate growth
only. Occasionally faint brownish pigment
produced.
Glucose-asparagine-meat
Growth hyaline, changing to orange-yellow
Aerial mycelium, if
extract agar:
or purplish-brown.
present, white, changing to ash-gray or dark
gray with tawny reverse. Faint yellowish
soluble pigment occasionally discernible.
Nutrient agar: Growth good, translucent
to brownish. No aerial mycelium. No soluble
pigment. Melanin-negative.
Potato: Growth lichenoid, light orange-
yellow to brown-red to purplish. No aerial
mycelium. Color of plug unchanged.
Gelatin: Cream-colored surface ring. Liq-
uefaction none to limited. No soluble pig-
ment.
Milk: Growth limited, yellow-brown sur-
face. Coagulation and peptonization variable
(often none, occasionally present).
Production of HS: Mostly negative.
Antagonistic properties: Produces chlor-
amphoteric compound
and
various
viruses.
tetracycline, an
both
chlorine, active against
rickettsiae, the
organism also produces, especially in a
chlorine-poor medium, tetracycline. The
presence of phosphorus in the medium in-
fluences not only growth but also antibiotic
containing nitrogen non-ionic
bacteria,
The
and larger
production (Prokofieva-Belgovskaya and
Popova, 1959).
Habitat: Soil.
Remarks: The numerous natural and
induced variants of S. aureofaciens display
wide variations in color of substrate growth,
ranging from pale yellow to reddish-brown,
and even occasionally greenish, depending
upon the composition of the nutrient sub-
strates and environmental conditions (Dug-
gar et al., 1954). Color of aerial mycelium
is influenced by sporulation. Ettlinger e¢ al.
(1958) included S. ambofaciens in this group.
Type culture: IMRU 3550; ATCC 10,762.
26. Streptomyces aureus (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, 8S. A. and Curtis, R. E. Soil Sci.
1: 24, 1916; 8: 97, 1919).
Morphology: Aerial
sporophores with numerous closed spirals;
mycelium forms
some strains produce flexible sporophores
with open spirals. Spores spherical to oval,
0.6, to 1.0 by 0.8 to 1.4 uw (Fig. 32).
Sucrose nitrate agar: Growth thin, spread-
ing, colorless, becoming dark brown. Aerial
mycelium thin, powdery, mouse-gray, be-
coming cinnamon-drab. No soluble pigment.
Growth cream-
black. Aerial
mycelium light brown. No soluble pigment.
Malate-glycerol
agar:
colored, with surface almost
178
Glucose-asparagine agar: Growth light
orange; raised center, hyaline margin. Aerial
mycelium hght drab.
Nutrient agar: Growth restricted, gray.
No aerial mycelium. Soluble pigment deep
brown.
Starch agar: Growth thin, transparent,
,
THE ACTINOMYCETES, Vol. II
Aerial buff-colored.
Good hydrolysis.
Potato: Growth abundant, wrinkled,
brown, becoming black. Aerial mycelium
white to ash-gray. Soluble pigment black.
Gelatin: Surface growth fair, cream-col-
Aerial mycelium
spreading. mycelium
ored, becoming brown.
Figure 32. Sporophores of S. aureus, X 30,000 (Courtesy of KE. Baldacci, University of Milan, Italy).
DESCRIPTION OF SPECIES OF STREPTOMYCES
absent or white. Brown soluble pigment.
Liquefaction rapid, later slowing down.
Milk: Black ring. Limited coagulation and
peptonization.
Nitrate: Reduction to nitrite with certain
‘carbon sources.
Invertase: None to positive.
Temperature: Optimum 25°C.
Antagonistic properties: Produces poly-
enes, substances active against various
fungi. Some strains produce luteomycin.
Habitat: Soil.
temarks: Cultures under this name were
described by DuBois-Severin in 1895, by
Lachner-Sandoval in 1899, and by Sartory
in 1923. Yamaguchi and Saburi (1955) re-
ported that the S. aureus culture obtained
from collections produces straight aerial
hyphae and no spirals when grown on var-
ious synthetic media. Okami and Suzuki
(1958) isolated two strains that produced
spirals. Ettlinger et al. (1958) considered
this organism as a strain of S. griseus. IKras-
silnikov (1949) considered it as a variety of
A. flavus.
Type culture: IMRU 3309.
27. Streptomyces autotrophicus Takamiya
and Tubaki, 1956 (Takamiya, A., and
Tubaki, K. Arch. Mikrobiol. 25: 58-64,
1956).
Morphology: Sporophores alternately or
irregularly branched, breaking up into
spores; no spiral formation. Spores colorless
with smooth surface; varying in shape from
ellipsoid to long ovoid or cylindrical; usually
2.5 to 4.8 by 0.5 to 0.8 uw, sometimes smaller,
ib by 0:3 %u-
Nitrate, carbohydrate-free, agar: Aerial
mycelium powdery and snow-white in ap-
pearance, consisting of a tough mycelial felt;
thicker at central area than at periphery.
Reverse side of growth wrinkled and pale
yellowish. No soluble pigment.
Calcium malate agar: Growth much
folded, and raised in central area, cream-
yellow at earlier stages of development;
reverse side pale brownish. Aerial mycelium
white.
Nutrient agar:
raised; reverse side relatively smooth
Growth much folded and
and
pale brownish. Aerial mycelium snow-white.
In old cultures, a faint brown tint in agar
layer immediately beneath growing colony.
Malt agar: Growth irregularly wrinkled
and folded; reverse side wrinkled and pale
yellowish. Production of spores rather poor.
Starch agar: Growth scanty; no hydroly-
sis.
Potato: Growth colorless, much folded,
with thick central area and thin periphery,
pale brownish.
Gelatin: No liquefaction.
Milk: Thin white pellicle formed on sur-
face. Reverse side yellowish. No coagulation.
No soluble pigment.
Cellulose: Not decomposed.
Nitrate reduction: None.
Habitat: Originally found on the surface
of phosphate buffer solution left unused in a
laboratory in Tokyo. Conceivably, it was
derived from atmospheric dust.
temarks: Hirsch (1960) considers this or-
ganism as a Nocardia (N. autotrophica), ca-
pable of utilizing petroleum.
28. Streptomyces beddardii (Erikson, 1935)
Waksman (Erikson, D. Med. Research
Council (Brit.) Spec. Rept. Ser. 203: 13-14,
1935).
Morphology: Sporophores long, slender,
forming many wavy or closely coiled spirals,
particularly on glucose agar; spirals less
marked or lacking on poorer nutritive media
like synthetic glycerol agar or water agar.
Aerial hyphae straighter and more branched
with shorter sporophores on starch agar.
Spores oval.
Glucose-asparagine agar: Growth wrin-
kled, membranous. Aerial mycelium scant,
white.
Nutrient agar: Growth colorless, coherent,
wrinkled, membranous. Aerial mycelium
scant, white. Soluble pigment deep brown.
180
Starch agar: Growth spreading, colorless.
Aerial mycelium abundant, white. Hydroly-
sis.
Egg medium: Growth extensive, wrinkled,
bright yellow. Considerable liquefaction.
Blood agar: Growth in uniformly striated
colorless bands; occasional round colonies at
margin. Hemolysis positive.
Potato: Growth moist, colorless. Aerial
mycelium scant, white at top of plug.
Gelatin: Dull white flakes sinking to bot-
tom as medium liquefies. Rapid liquefaction.
Milk: Coagulation followed by peptoniza-
tion.
Source: Human spleen in a case of splenic
anemia. No record concerning actual patho-
genicity.
28a. Streptomyces bellus Margalith and
Beretta (Margalith, P. and Beretta, G.
Mycopathol. Mycol. Appl. 12: 189-195,
1960).
Morphology :
flexuous with short, open hooks.
Sucrose nitrate agar: Substrate growth
light cherry-pink. Aerial mycelium pinkish-
white. Soluble pigment light pink.
Glucose-asparagine agar: Growth colorless
to hight pink with orange tinge. Aerial myce-
lium white, with small amount of bluish
spores. Soluble pigment lght pink with
Sporophores long, straight,
orange tinge.
Calcium malate agar: Substrate growth
pink, with pinkish-violet reverse. Sporula-
tion abundant, pinkish-blue. Soluble pig-
ment pinkish-violet.
Nutrient agar: Substrate growth hyaline
with Aerial mycelium
slight or absent. Soluble pigment brown.
Starch agar: Growth hyaline with color-
less to pale pink reverse. Aerial mycelium
brownish reverse.
limited, with bluish-green spores. Starch
hydrolyzed.
Glucose-casein — digest-yeast-beef
Growth pink. Aerial mycelium pinkish with
bluish-green spores. Soluble pigment light
agar:
pink.
THE ACTINOMYCETES, Vol. II
Glucose-yeast extract-beef-peptone agar:
Growth pink. Aerial mycelium pinkish-
white with bluish-green spores. Soluble pig-
ment reddish-brown.
Potato: Growth rough, colorless to brown.
No aerial mycelium. Soluble pigment brown.
Gelatin: Slow liquefaction. Soluble pig-
ment brown.
Milk: Growth in form of brownish ring.
Nitrate reduction: None.
Cellulose: Moderate growth.
Carbon utilization: Most sugars readily
utilized. Xylose, inulin, and dulcitol not
utilized in solid media. Succinate, citrate and
glycine not utilized.
Antagonistic properties: Produces anti-
biotic matamycin, active upon gram-positive
bacteria.
Habitat: Soil in Italy.
29. Streptomyces bikiniensis Johnstone
and Waksman, 1948 (Johnstone, D. B. and
Waksman, 8. A. J. Bacteriol. 55: 317-326,
1948).
Morphology: Sporophores straight. Spores
oval (Fig. 33).
Sucrose nitrate agar: Growth white, be-
coming pallid neutral gray with white tinge.
Aerial mycelium abundant, white to gray.
Soluble pigment light brown. Superficial
droplets amber-colored.
Glucose-asparagine agar: Growth abund-
ant. Aerial mycelium white to mouse-gray.
Soluble pigment light amber.
Nutrient agar: Growth luxuriant. Aerial
mycelium moderate, white. Soluble pigment
deep brown.
Starch agar:
mycelium white, becoming gray. Slight hy-
Growth abundant. Aerial
drolysis.
Potato: Growth wrinkled and raised, pale
ochraceous buff. Soluble pigment brown to
black.
Gelatin: Shght liquefaction.
Milk: Surface growth patchy, white.
Aerial mycelium gray. Gradual peptoniza-
tion.
DESCRIPTION OF
ay
4i2@
S. bikiniensis,
FIGURE 33.
tesy of kK. L. Jones).
Production of HoS: Positive.
Strongly
Produces streptomycin.
Source: Soil from Bikini Atoll.
Type culture: IMRU 3514
Antagonistic properties: antago-
nistic.
30. Streptomyces blastmyceticus Watanabe
et al., 1957 (Watanabe, K., Tanaka, T.
Fukuhara, K., Miyairi, N., Yonehara, H..,
and Umezawa, H. J. Antibiotics (Japan)
1OA: 39-45, 1957).
SPECIES OF
Ky
erown on casein digest-beef extract
STREPTOMYCES LS]
aa”
es
lee
/
agar for 12 days, X 3,000-4,000 (Cou1
Morphology: Sporophores straight. Spores
oval to spherical, 1 by 1.5 u.
Sucrose nitrate agar: Growth weak, color-
less or white. Aerial mycelium poor, white.
No soluble pigment.
Glucose-asparagine agar: Growth good,
=
colorless, later cream-colored. No aerial my-
celium. No soluble pigment.
Calcium citrate-glycerol agar: Growth
colorless or white, later deep olive-buft.
182
Aerial mycelium thin, powdery, white or
pale yellow to pale olive-buff.
Nutrient agar: Growth white, later cream-
colored to light brown. Aerial mycelium
poor, white to gray. Soluble pigment brown.
Milk: Growth in the form of ring on sur-
face, cream-colored to brown. Aerial myce-
lium white. Soluble pigment brown. Rapid
peptonization.
Potato plug: Growth gray to olive-gray.
Aerial mycelium white. Usually no soluble
pigment.
Nitrate reduction: Negative.
Starch: Hydrolyzed.
Carbon utilization: Utilizes glucose, fruc-
tose, galactose, or starch. Grows poorly on
sucrose, lactose, maltose, or inositol. Does
not utilize xylose, arabinose, raffinose, rham-
nose, mannitol, sorbitol, dulcitol, or salicin.
Antagonistic properties: Produces an
antifungal agent designated as blastmycin.
Remarks: Related to S. flavochromogenes.
31. Streptomyces bobiliae (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, 8S. A. and Curtis, R. E. Soil Sci.
1: 121, 1916; 8: 100, 1919).
Morphology: Elongated sporophores form
a few close spirals of a dextrorse type. No
spirals according to Jensen (1930). Spores
oval and spherical.
Sucrose nitrate agar: Growth abundant,
wrinkled, coral-red becoming deep_ red.
Aerial mycelium scant, white; later absent.
No soluble pigment.
Glycerol malate agar: Growth cinnamon-
buff. No aerial mycelium.
Glucose-asparagine agar: Growth coral-
red. No aerial mycelium.
Nutrient agar: Growth gray, becoming
brownish to coral-red. No aerial mycelium.
Soluble brown pigment in presence of glyc-
erol (Jensen).
Potato: Growth thin, dry, and wrinkled,
yellowish, becoming coral-red. No aerial my-
celium. Soluble pigment grayish to black.
Gelatin: Growth cream-colored to orange.
THE ACTINOMYCETEHS, Vol. II
Aerial mycelium in the form of occasional
patches of white. Rapid liquefaction. Soluble
pigment brown. Melanin-positive.
Milk: Dark brown ring. Peptonization
without coagulation.
Starch media: Growth wrinkled, coral-red
with hyaline margin. Aerial mycelium white.
Hydrolysis medium.
Nitrate: Good reduction to nitrite.
Cellulose: No growth in solution. Good
growth on plate.
Invertase: Positive.
Production of H.S: Positive.
Temperature: Optimum 37°C.
Antagonistic properties: Produces
mented antibiotic cinerubin.
Habitat: Common in soil.
Remarks: S. purpurascens is considered by
Corbaz et al. (1957) as a synonym of S.
bobiliae, except that the latter no longer
produces any aerial mycelium or spores.
IKxrassilnikov (1949) considered this species
pig-
as a variety of A. ruber.
Type culture: IMRU 3310.
32. Streptomyces bottropensis — IKonink.
Nederl. Gist et Spirit. (IXonink. Nederland.
Gist et Spirit. Brit. Pat. 762,736, Dec._5,
1956).
Morphology: Aerial mycelium ramified,
with short, open spirals. Spores cylindrical,
elliptical to almost spherical, | to 4 by 0.6
LowlZau:
Sucrose nitrate agar: Growth abundant,
reddish. Aerial mycelium limited. Soluble
pigment brown.
Glucose-asparagine agar: Growth good,
yellow. Aerial mycelium limited, white to
pale gray. No soluble pigment.
Calcium malate agar: Growth good, vel-
lowish-brown. Aerial mycelium white-gray.
No soluble pigment.
Starch agar: Growth at first pink, later
darker (pH sensitive; acid-pink, alkaline
blue). Aerial mycelium limited, white to
gray. Starch hydrolyzed.
Glucose
nutrient agar: Growth folded,
DESCRIPTION OF SPECIES OF STREPTOMYCES
yellowish. Aerial mycelium abundant, white.
No soluble pigment.
Glucose-yeast extract-peptone — agar:
Growth yellowish to buff. Aerial mycelium
white to gray. No soluble pigment.
Potato agar: Growth smooth, yellowish-
brown. Aerial mycelium white to bluish-
eray. Soluble pigment at first absent, later
dark.
Gelatin: Growth on surface good. Ae. ial
mycelium white. Rapid liquefaction. Soluble
pigment dark brown.
Potato: Growth folded, brown to black.
No aerial mycelium.
Milk: Growth moderate. No coagulation;
no peptonization.
Antagonistic properties:
biotic B-mycin, active against cocci, gram-
positive bacteria, and mycobacteria.
Habitat: Soil.
Remarks: This is one of the organisms
that can be either melanin-negative (nutri-
ent agar, yeast extract agar) or melanin-
positive (gelatin, potato agar).
Produces anti-
33. Streptomyces brasiliensis (Spencer,
1921) Waksman (Spencer, E. R. Botan.
Gaz. 72: 285-287, 1921).
Morphology: Aerial mycelium forms
straight, branched sporophores. Spores borne
in chains on free ends of hyphae, oblong,
1.6. by 0.8 yu.
Sucrose nitrate agar: Growth at first
white; after 10 days pale pinkish-buff. Aerial
mycelium white and dense. No soluble pig-
ment.
Glucose-asparagine agar: Growth luxuri-
ant, color same as on sucrose nitrate agar,
thallus conspicuously zonated. Aerial myce-
lium powdery, white to pale pinkish-buff. No
soluble pigment.
Glycerol malate agar: Growth spreading
and not zonated, bordered by submerged
mycelial bands of varying width, pearl-
white. Aerial mycelium short, loose, and
pearl-white.
Potato: Growth vigorous, crumpled, pale
183
pinkish-buff. Aerial mycelium abundant, at
first white, later pale pinkish-buff. No solu-
ble pigment. Melanin-negative.
Nut plugs: Growth vigorous, pale pinkish-
buff. Aerial mycelium powdery, white.
Medium not completely but
much shrunken and blackened.
Gelatin: Rapid liquefaction. No soluble
pigment.
Milk: Rapid coagulation and peptoniza-
destroyed,
tion.
Habitat:
nuts.
Remarks: This Streptomyces species is to
be distinguished from Nocardia brasiliensis,
Parasitic on kernels of Brazil
a pathogenic organism.
34. Streptomyces cacaoi (Waksman, 1932)
Waksman and Henrici, 1948 (Waksman, S.
A. In Bunting, R. H. Ann. Appl. Biol. 19:
515-517, 1932).
Morphology: Sporophores long; spirals
long and open, not compact.
Sucrose nitrate agar: Growth thin, yel-
lowish, later turning reddish-brown. Aerial
mycelium light gray to mouse-gray, with
white edge. No soluble pigment.
Nutrient agar: Growth brown, covered
with tiny patches of ivory-colored aerial
mycelium.
Potato: Growth abundant, brownish.
Aerial mycelium white to mouse-gray. Mel-
anin-negative.
Gelatin: Growth No aerial
mycelium. Liquefaction rapid. No soluble
flocculent.
pigment.
Nitrate reduction: Limited.
Production of HS: Negative.
Antagonistic properties: Certain strains
produce an antibiotic designated as cacao-
mycetin.
Source: Three strains were isolated from
cacao beans in Nigeria. They showed slight
differences, the foregoing description being
based on one strain.
Remarks:
strong diastatic action, no sugar or dextrin
Strong proteolytic enzymes,
184
being left in 1 per cent starch solution after
a few days.
Type culture: IMRU 3082.
35. Streptomyces caelestis DeBoer et al.,
1959 (DeBoer, C., Dietz, A., and Hoeksema,
H. Canad. Pat. 572,779, March 24, 1959).
Morphology: Sporophores loosely coiled.
Spores spherical to oval.
Sucrose nitrate agar: Growth good. Aerial
mycelium gray-white. Soluble pigment yel-
low.
Nutrient agar: Growth fair to good. Aerial
mycelium slight pink-white. Soluble pigment
brown-tan.
Casein digest-beef agar:
Aerial mycelium pale glaucous blue. Soluble
pigment brown-tan.
Starch agar: No growth.
Growth good.
Tyrosine agar: No growth. No aerial my-
celium. No soluble pigment.
Potato: Growth good. Aerial mycelium
grayish to blue-white. Soluble pigment
brown.
Gelatin: Growth good. Aerial mycelium
blue-gray. Soluble pigment brown. Medium
liquefaction.
Milk: Growth fair. No soluble pigment.
No peptonization.
Nitrate reduction: Negative.
Production of H.oS: Positive.
Carbon utilization: Utilizes a variety of
sugars, dl-inositol, acetate; limited utiliza-
tion of starch, glycerol, citrate, and succi-
nate; utilize duleitol, mannitol,
inulin, sorbitol, and various other organic
does not
acids.
Antagonistic properties: Produces antibi-
otic celesticetin.
Habitat: Soil in Utah.
Remarks: Similar to S. glaucus and SN.
chartreusis.
36. Streptomyces caeruleus — (Baldace1)
Waksman (Baldacci, E. Atti ist.
univ. Pavia 3: 180-184, 1944).
Morphology: Sporophores long, straight,
botan.
THE ACTINOMYCETES, Vol. II
branched, not forming any spirals. Spores
eylindrical 1.0 to. 1.4 by 2.0 toi2-) nu:
Agar media: Substrate growth colorless.
Aerial mycelium pigmented, at first white,
later becoming blue, and finally dark. Solu-
ble pigment grayish-green.
Glycerol agar: Grows slowly; light blue in
color.
Carrot agar: Growth at first white; later
becoming blue. Aerial mycelium blue, be-
coming gradually deep blue, and_ finally
dark blue.
Oatmeal agar: Color of growth at first
white and aerial mycelium blue, gradually
becoming darker in color. The agar is pig-
mented grayish-green.
Gelatin: Growth erayish-blue.
Either no liquefaction or only slow liquefac-
slow,
tion.
Milk: Weak growth.
Starch media: Weak greenish growth.
Bluish-green pigmentation.
Temperature: Range between 18 and
30°C. Optimum 24°C.
teaction: Optimum pH 8 to 10.
Cellulose: Not utilized.
Antagonistic properties: Produces anti-
biotic caerulomycin.
Habitat: Corn straw and decomposing
rice straw.
femarks: Related to S. violaceoruber, S.
violaceus, and S. viridis. According to Taber
(1959) the distinctive characteristics of a
culture that he isolated and identified as
S. caeruleus are: production of a blue to red
indicator pigment; requirement of a neutral
or alkaline reaction for growth and produc-
tion of oblong to cylindrical spores in straight
and flexuous chains. It was not chromogenic
on peptone media but produced HS on
iron-peptone agar. It did not grow on un-
buffered potato or carrot plugs, litmus
milk, and certain synthetic agar media. The
culture readily utilized glucose, fructose,
galactose, mannitol, sucrose, xylose, starch,
and maltose, but did not utilize, or utilized
DESCRIPTION OF SPECIES OF STREPTOMYCES
to a limited extent, mannose, ?7-Inositol,
adonitol, lactose, ribose, raffinose, and cel-
lulose.
Type culture: IMRU 3798.
37. Streptomyces caespitosus Sugawara and
Hata, 1956 (Sugawara, R. and Hata, T.
J. Antibiotics (Japan) 9A: 147-151, 1956).
Morphology: Primary verticils produced.
Spores oval, 1.3 to 0.5 by 0.3 to 0.5 u.
Sucrose nitrate agar: Growth hyaline,
colorless to faint yellowish-brown. Aerial
mycelium white to yellowish-gray to green-
ish-yellow. Soluble pigment faint yellow.
Calcium malate Growth
with yellow-brownish center, changing to
dark greenish-yellow to dull yellow. Aerial
yellowish
agar: colorless
mycelium white, with tinge,
greenish-yellow at the margin. Soluble pig-
ment faint yellow, pinkish shade in some
cultures.
Glucose-peptone agar: humid,
wrinkled, cracked in the center; colorless,
becoming greenish-yellow-gray, — reddish-
gray, to dark gray. Aerial mycelium thin,
gray. Soluble pigment reddish-brown.
Starch agar: Growth scanty, colorless to
Growth
faint yellowish-brown, or yellow to orange-
yellow. Aerial mycelium cottony, white,
cream with lavender patch in the center,
becoming yellowish-gray. Soluble pigment
faint yellowish-brown.
Nutrient agar: Growth yellow-brown to
gray to dark gray. Aerial mycelium gray.
Potato: Growth cream to brownish, center
light greenish-yellow. Aerial mycelium white
to grayish or gray with pale olive tinge.
Soluble pigment absent, or dark brown, or
grayish-brown.
Milk: Surface ring yellow to pale yel-
lowish-brown. No aerial mycelium. Soluble
pigment pale brown.
Gelatin: Growth cream-colored turning
greenish-yellow to reddish-yellow. Aerial
mycelium white to yellow. Soluble pigment
yellowish-brown. Rapid liquefaction.
Tyrosinase reaction: None.
LS5
Nitrate reduction: Positive.
Starch: Hydrolysis.
Carbon utilization: Utilizes various car-
bohydrates; does not utilize xylose, rham-
nose, raffinose, arabinose, mannitol, salicin,
dulcitol, inulin, acetate, and succinate.
Antagonistic properties: Produces anti-
biotic mitomycin, active upon certain
neoplasms.
temarks: Closely related to S. kitasatoen-
sis and S. hachijoensis.
38. Streptomyces caiusiae Dhala et al.,
1957 (Dhala, 8S. A., Poonawalla, F. M., and
Bhatnagar, 8S. S. J. Sci. & Ind. Res. 16C:
76-80, 1957).
Morphology: Aerial hyphae short and
straight; frequently clusters are produced,
subdividing at the distal portions into chains
of spores. No spirals formed either in syn-
thetic or nonsynthetic media; often tips of
the aerial hyphae slightly curved. Spores
round to oval, 0.6 to 1.4 by 0.4 to 0.8 yu.
nitrate agar: Colonies round,
convex, tough, with smooth surface when
unsporulated; citron-yellow, later turning
brown. Aerial mycelium white, turning yel-
lowish, then gray. Soluble pigment at first
yellow but later darkened to a brown tinge.
Glucose-asparagine agar: Growth citron-
vellow. Aerial mycelium white. Soluble pig-
Sucrose
ment yellowish-brown.
Nutrient agar: Growth
irregular margins and radial ridges in old
cultures. White aerial mycelium.
Starch agar:
after 2 days; on further incubation, they
wrinkled, with
Colonies smooth, colorless
become large and wrinkled with radiating
ridges. Aerial mycelium white, turning gray.
Soluble pigment brown. Starch weakly hy-
drolyzed.
Potato: Growth luxuriant, citron-yellow.
Aerial mycelium white, turning gray. Plug
turns black.
Milk: Pellicle produced. Coloration of
milk brownish to black. Peptonization posi-
tive.
186
Gelatin: Sediment buff-colored. Liquefac-
tion medium.
Tyrosinase reaction: Positive.
Nitrate reduction: None.
Carbon sources: Sugars readily utilized,
with the exception of acetate, benzoate,
cellulose, dulcitol, 2-inositol, and salicylate.
Antagonistic properties: Active primarily
upon gram-negative bacteria and fungi.
Remarks: Closely related to S. antzbi-
oticus.
39. Streptomyces californicus (Waksman
and Curtis, 1916) Waksman and Henrici
(Waksman, S. A. and Curtis, R. E. Soil Sci.
1: 22, 1916;\Waksman, 8. A. zbed. 8: 104,
1919).
Synonyms:
Streptomyces puniceus Finlay and Sobin,
1950.
Streptomyces vinaceus Mayer et al.,
1951.
Streptomyces floridae Bartz et al., 1951.
Streptomyces griseus var. purpureus
Burkholder e¢ al., 1955.
Streptomyces purpureus (Burkholder,
1955) Waksman, 1959.
Morphology: The original culture was re-
ported to form sporophores with long, nar-
row, open sinistrorse spirals. According to
Okami, however, the
straight. Recent examinations of the original
culture of Waksman and Curtis (Burkholder
et al., 1955) did not reveal any spirals either.
Sucrose nitrate agar: Growth spreading,
Aerial mycelium
powdery, light neutral gray to ash-gray. No
soluble pigment.
sporophores — are
vinaceous-colored.
Glucose-asparagine agar: Growth — re-
stricted, much folded, cream-colored, with
sulfur-yellow tinge.
Nutrient
yellowish to cream-colored, Melanin-nega-
agar: Growth thin, restricted,
tive.
Starch Growth
center with colorless to gray margin. Hy-
spreading, pink
agar:
drolysis rapid.
THE ACTINOMYCETES, Vol. II
Potato: Growth glossy, yellow to red,
turning red-brown.
Gelatin: Growth gray, moist, abundant.
No soluble pigment. Liquefaction medium.
Milk: Surface growth faint, brownish.
Coagulation and slow peptonization.
Nitrate reduction: Positive.
Production of H.S: Negative.
Cellulose: Growth scant but definite.
Carbon utilization: According to Burk-
holder, the various strains utilize p-xylose,
b-glucose, p-galactose, p-fructose, cellobiose,
p-maltose, p-mannitol, and starch. Growth
poor with L-arabinose, L-rhamnose, p-lac-
tose, sucrose, D-raffinose, dulcitol, 2-inositol,
and salicin.
Temperature: Optimum 37°C.
Invertase: Positive.
Antagonistic properties: Routien and
Hofmann (1951) first demonstrated that
cultures of S. californicus are capable of
producing viomycin. The same antibiotic
was found to be produced by other strains
of this organism.
Habitat: Soil.
temarks: Waksman and Curtis reported
the production of spirals in their original
description of S. californicus (see also Waks-
man, 1919). Several authors who studied
the S. californicus culture more recently de-
scribed the aerial mycelium as straight to
wavy to strongly flexuous (Burkholder et al.,
1955; Kutzner, 1956; Ettlinger et al., 1958).
Since the other properties of the now-avail-
able S. californicus fit with the original de-
scription, it might be assumed that the
strongly flexuous aerial hyphae were con-
sidered as spirals originally. In 1955 Burk-
holder et al. made a comparative study of
several viomycin-producing organisms which
were originally described under the names
S. floridae, S. puniceus, S. vinaceus, S. calr-
fornicus, and several others. All these cul-
tures behaved in a similar manner, with
only minor differences between this group
and S. californicus ATCC 3312; namely,
DESCRIPTION OF SPECIES OF STREPTOMYCES 187
“with the exception of ATCC 3312, all
isolates liquefy gelatin rapidly and produce
viomycin or similar antibiotic compounds.”
No studies seem to have been made, how-
ever, of antibiotics produced by S. californi-
cus ATCC 3312. Only because the original
description of S. californicus gave spiral for-
mation and these organisms did not, the
several viomycin-producing or
the S. californicus ATCC 33
scribed as a variety of S. griseus, namely
S. griseus var. purpureus. This was due to
the fact that S. griseus had the same mor-
ganisms and
12 were de-
phology and color of the aerial mycelium,
and because several streptomycin-producing
strains are known to form also a red-gray
color in the substrate growth.
Type culture: IMRU (ATCC) 3312.
40. Streptomyces calvus Backus et al.,
1957 (Backus, E. J., Tresner, H. D., and
Campbell, T. H. Antibiotics & Chemother-
apy 7: 532-541, 1957).
Morphology: Sporophores form
loose spirals. Spores globose to elongated,
0.6 to 1.0 by 1.0 to 1.8 » (Fig. 34).
Sucrose nitrate agar: Growth cream-col-
short
ored to yellowish. Aerial mycelium scanty,
white to gray.
Glucose-asparagine agar: Growth ivory-
yellow. Aerial mycelium scanty, white.
Calcium malate agar: Growth colorless to
vellow. Aerial mycelium scanty white to
gray. Crystalline pellets formed in growth
ZONES.
Starch agar: Growth colorless to yellowish.
Aerial mycelium white to mouse-gray.
Nutrient agar: Substrate growth light yvel-
low. Aerial mycelium scanty, white. No
soluble pigment. Melanin-negative.
Potato plug: Growth gray. Aerial myce-
hum scanty, white to light gray. Plug dis-
colored.
Gelatin: Growth colorless to yellow. No
aerial mycelium. Partial liquefaction. No
soluble pigment.
Milk: Growth colorless to yellow. Coagu-
lation and moderate peptonization.
Cellulose: Growth yellow. No decomposi-
tion of cellulose.
Production of HoS: Negative.
Carbon utilization: d-fructose, 7-inositol,
lactose, d-mannitol, d-raffinose, /-rhamnose,
sucrose, d-trehalose, and d(+-)-xylose readily
utilized; /-arabinose, d-melibiose, and_ sali-
cin utilized poorly; dextrin, esculin, deme-
lezitose, and adonitol not utilized at all.
Antagonistic properties: Produces
nucleocidin, an antibiotic possessing anti-
trypanosomal properties.
Habitat: Soil in India.
Remarks: This organism is closely related
to S. annulatus.
41. Streptomyces candidus (Krassilnikov,
1941) Waksman (Not Streptothrix candida
Petruschky). CUsrassilnikov, N. A. Actino-
mycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 49, 1941).
Morphology: Sporophores long, straight
or wavy, but never forming spirals; occa-
sionally arranged in broom-shaped_ bodies
or fascicles. Spores oblong to cylindrical
(‘fragmentation spores”), 1.0 to 2.0 by
0.6 to 0.8 x.
Sucrose nitrate
Aerial mycelium velvety, white. No soluble
agar: Growth colorless.
pigment.
Nutrient agar: Growth good, lichenoid
or smooth. Aerial mycelium whitish. Mela-
nin-negative.
Gelatin: Slow liquefaction. Melanin-nega-
tive.
Potato: Growth colorless, lichenoid. Aerial
mycelium poorly developed. No soluble pig-
ment or brownish.
Milk: No coagulation; good peptoniza-
tion.
Starch: Rapid hydrolysis.
Cellulose: Good growth.
Nitrate reduction: Positive.
Sucrose: Inversion.
Production of HS: Negative.
188
THE ACTINOMYCETES, Vol. II
FriGcure 34. Sporophores of S. caluus (Reproduced from: Backus, E. J. et al. Antibiotics & Chemo-
therapy 7: 536, 1957).
Antagonistic properties: Weak.
Habitat: Soil.
Remarks: Certain varieties of this species
have also been described. This is true, for
example, of A. fasciculus, which Krassilni-
kov himself considered as a variety of S.
candidus; it 1s true of A. farinosus
Krassilnikov and of A. candidus var. albo-
roseus described by Gause et al. (1957).
S. nitrosporeus Okami (1952) appears to be
closely related, if not identical to it. Ett-
linger et al. (1958) consider this organism
also
as related to S. griseus.
Type culture: IMRU 3416.
42. Streptomyces canescus Hickey et al.,
1952, (Hickey, “Ri J.;- Corum, ©. J.) "midy,
P, H., Cohen; I.R:; Nagers U.F= Band
Kropp, E. Antibiotics & Chemotherapy
2: 472-483, 1952).
Morphology: Sporophores straight — or
curved, not forming any spirals. Spores
globose, 1.0 to 1.3 by 1.3 to 2.6 p (ig. 35)
Calcium malate agar: Growth gray to
rose-gray; reverse yellow to tan. No soluble
pigment.
Yeast extract-casein digest agar: Growth
effuse to convex, edge filamentous; reverse
brown. Aerial mycelium powdery, varying
from gray-white to gray. No soluble pig-
ment.
Acid-glucose-peptone Growth at
first white, then tan. Aerial mycelium faintly
agar:
z
DESCRIPTION OF SPECIES OF STREPTOMYCES LS9
greenish, produced after 14 days. Amber Milk: Soft, rennet curd formed at 36°C
pigment diffused throughout medium. after 48 hours; completely peptonized in 12
Kee medium: Growth tan, wrinkled. No days.
sporulation after 10 days; limited white Starch: Hydrolysis strong.
sporulation observed in 14 days. Soluble Nitrate reduction: Negative.
pigment brown. Very slow liquefaction after Production of H.S: Negative.
28 days. Temperature: Optimum 36°C
Potato: Growth lhght gray, wrinkled. Carbon utilization: Utilizes glucose, arabi-
Soluble pigment deep brown. nose, trehalose, xylose, sucrose, maltose,
Gelatin: Liquefaction rapid. Soluble pig- galactose, dextrin, soluble starch, mannitol,
ment deep brown. glycerol, and salicin. No growth with sor-
¥
sl], A :
Fiat RE 35. S. canesc us, grown on casein digest beef extract agar for 12 days Xx 3.000—4.000 (Courtesy
Le
@
of kK. L. Jones
190
bose, melezitose, dulcitol, rhamnose, sorbi-
tol, melibiose, phenol, raffinose, and lactose.
Antagonistic properties: Produces anti-
fungal antibiotic ascosin.
Source: Contaminated fungus plate.
Remarks: This species is now included
with S. coelicolor (Kutzner and Waksman,
1959).
Type culture: IMRU 3782; NRRL 2419.
43. Streptomyces canus Heinemann et al.,
1953 (Heinemann, B., Kaplan, M. A., Muir,
R: D:..and> Hooper. i... Antibiotics: .&
Chemotherapy 3: 1239-1242, 1953).
Morphology: Aerial mycelium forms nu-
merous loosely wound spirals. Spores spher-
oidale 0 toa..2 by 1:6 towl.8: a:
Sucrose nitrate agar: Growth moderate,
wrinkled, yellow-brown. Aerial mycelium
secant. No soluble pigment.
Glycerol-asparagine agar: Growth abun-
dant, cream-colored, turning russet-brown
with aging. Aerial mycelium abundant,
slate-gray. Soluble pigment amber.
Calcium malate agar: Growth moderate,
golden colored. Aerial mycelium scant. No
soluble pigment.
Nutrient agar: Growth abundant, yellow.
Aerial mycelium white to light yellow. Solu-
ble pigment faint yellow.
Potato: Growth abundant, cream-colored.
No aerial mycelium. Slight reddish-brown
darkening of the potato. Melanin-negative.
Gelatin: Moderate liquefaction at 26°C
in 14 days. No soluble pigment.
Milk: Alkaline with no coagulation; slight
peptonization in 14 days.
Starch: Hydrolysis in 96 hours at 30°C.
Nitrate reduction: Positive in 96 hours
at 30°C.
Carbon utilization: Good growth with
arabinose, rhamnose, xylose, dextrose, galac-
tose, fructose, cellobiose, lactose, maltose,
sucrose, dextrin, inulin, raffinose, soluble
starch, glycerol, inositol, mannitol, and
sodium salicylate. No growth observed with
dulcitol, sorbitol, sodium acetate, sodium
THE ACTINOMYCETES, Vol. II
citrate, sodium formate, sodium malate,
sodium oxalate, sodium tartrate, or sodium
succinate.
Antagonistic properties: Produces the
antibiotic amphomycin, active against gram-
positive bacteria.
Habitat: Soil.
Type culture: ATCC 12,237.
44. Streptomyces carnosus (Millard and
Burr, 1926) Waksman and Henrici (Millard,
W. A. and Burr, S. Ann. Appl. Biol. 13:
580, 1926).
Morphology: Spores cylindrical, 1.0 by
OFT amis.
Sucrose nitrate agar: Growth pale smoky-
gray to olive-gray. Aerial mycelium abun-
dant, gray. Colorless guttation drops appear
over the whole surface. Soluble pigment
ivory-yellow to cartridge-buff.
Potato: Growth lichenoid. Aerial
celium gray to brownish with white spots.
Plug becomes colored gray to black.
Gelatin: Surface growth. Aerial mycelium
white in center, gray at margin. Rapid lique-
faction. Soluble brown pigment.
Milk: Surface growth good. No aerial my-
celium. Positive coagulation and peptoniza-
tion.
Starch: Hydrolysis.
Tyrosinase reaction: Negative.
Nitrate reduction: Positive.
Habitat: Potato scab.
my-
45. Streptomyces catenulae Davisson and
Finlay, 1959 (Davisson, J. W. and Finlay,
A. C. U.S. Pat. 2,895,876, July 21, 1959).
Morphology: Growth with
smooth edge. Sporophores in form of short
wrinkled
clusters; a few tight spirals. Spores oval to
cylindrical, 1.0 by 1.3 wu.
Agar media: Growth dark brown to dark
greenish-brown. Aerial mycelium dark olive-
gray to brown.
Sucrose nitrate agar: Growth transparent,
white. Aerial mycelium light gray. No solu-
ble pigment.
DESCRIPTION OF SPECIES OF STREPTOMYCES 19]
Calcium malate agar: Growth poor. Aerial
mycelium mouse-gray, with some white. Cal-
cium malate digested.
Nutrient agar: Growth pale yellow. Aerial
mycelium white, turning pale gray. Soluble
pigment pale yellow. Nonchromogenic.
Glucose-yeast extract-beef-peptone agar:
Growth dark brown. Aerial mycelium olive-
gray with white. Soluble pigment medium
brown.
Starch agar: Growth
Aerial mycelium poor, mouse-gray. Good
hydrolysis.
Gelatin: Growth moderate. Aerial myce-
lium buff, some white. No soluble pigment.
Poor liquefaction.
Potato: Growth good, greenish. Aerial my-
celium pale olive to smoke-gray to brown.
Soluble pigment dark greenish or absent.
Nitrate reduction: None.
Antagonistic properties: Produces
biotic catenulin.
Habitat: Soil.
Type culture: ATCC 12,476.
46. Streptomyces cavourensis Giolitti, 1958
(Giolitti, G. Belgian Pat. 560,930, March 18,
1958*).
Morphology: Aerial mycelium produces
spirals on certain media. Spores spherical to
elliptical.
Sucrose nitrate agar: Growth yellowish.
Aerial mycelium chalky white to yellowish.
brownish-orange.
anti-
No soluble pigment.
Glycerol-asparagine agar: Growth hazel-
colored. Aerial mycelium whitish. Soluble
pigment faint brown.
Calctum malate agar: Growth scanty,
yellow-brownish. Aerial mycelium scanty,
white. Soluble pigment scarce, yellow-brown-
ish.
Nutrient
Aerial mycelium scanty, chalky white to yel-
agar: Growth orange-brown.
lowish. Soluble pigment light brown.
Glucose agar: Growth brown, wrinkled.
* Supplemented by personal communication.
Aerial mycelium white-yellowish. Soluble
pigment dark brown.
Potato agar: Growth dark brown. Aerial
mycelium gray with dark yellow dots. Solu-
ble pigment brown.
light brown,
wrinkled. Aerial mycelium gray with brown
patches. Soluble pigment light brown.
Starch agar:
brownish. Aerial mycelium scanty, white-
grayish. Soluble pigment brownish. Strong
hydrolysis.
Gelatin: Growth scanty, brown, wrinkled.
Aerial mycelium scanty, gray. Soluble pig-
Oatmeal Growth
agar:
Growth scanty, yellow-
ment brownish. Liquefaction fairly good.
Potato: Growth brown with yellow edges,
wrinkled. Aerial mycelium grayish. Soluble
pigment brown.
Milk: Growth consists of white to yellow
ring around surface. Positive coagulation and
peptonization.
Nitrate reduction: Negative.
Antagonistic properties: Produces flaven-
somycin, an antibiotic active against fila-
mentous and yeast-like fungi, and to a
certain extent some gram-positive bacteria.
Very active against some insects.
Type culture: IMRU 3758.
47. Streptomyces celluloflavus Nishimura
et al., 1953 (Nishimura, H., Kimura, T., and
Kuroya, M. J. Antibiotics (Japan) 6A: 57—
65, 1953).
Morphology: Sporophores straight with a
few flexible, hooked spirals. Spores nearly
spherical, 1.0 by 0.9 x.
Sucrose nitrate agar: Growth glossy, de-
veloping deep into medium, later becoming
yellow. Soluble pigment faint sulfur-yellow.
malate agar: Growth yellow,
Glycerol
later turning white to pale olive-buff with
blackish center. Aerial mycelium cottony,
white, with grayish patches, later turning
olive-buff. Soluble pigment yellow.
Glucose-asparagine agar: Growth cream
to yellow. Aerial mycelium scant, cottony,
192 THE ACTINOMYCETES, Vol. II
white to gray. Soluble pigment sulfur-yel-
low.
Nutrient agar: Growth olive-buff, turn-
ing colorless. Aerial mycehum scant, cot-
tony, white to grayish. Soluble pigment
yellow with tinge of green to gold.
Potato: Growth wrinkled, deep olive-buff.
Aerial mycelium white to olive-buff. Soluble
pigment deep olive-buff.
Gelatin: Growth ivory-yellow to olive-buff
on surface of liquefied layer. No aerial my-
celium. Faint brownish pigment. Rapid to
medium liquefaction.
Milk: Growth yellow to dark olive-buff.
Aerial mycelium white. Soluble pigment red-
dish-brown. Coagulation and rapid peptoni-
zation.
Tyrosine medium: Growth ivory-yellow to
cream-buff. Aerial mycelium absent or scant
white. Soluble pigment greenish-yellow.
Cellulose agar: Growth poor. Soluble pig-
ment yellow.
Production of HS: Negative.
Antagonistic properties: Produces thio-
lutin, aureothricin.
Habitat: Soil.
48. Streptomyces cellulosae — (IXrainsky,
1914) Waksman and Henrici, 1948 (x<rain-
sky, A. Centr. Bakteriol. Parasitenk. Abt.
IT., 41: 683-688, 1914).
Description after Jensen, H. L. Soil Sei.
30: 65, 1930.
Morphology:
spiral formation. Spores almost spherical, 1.5
Sporophores straight; no
uw in diameter.
Sucrose nitrate agar: Growth at first trans-
parent, becoming lemon-yellow. Aerial my-
celium light gray, later deep slate-gray. Sol-
uble pigment may be lemon-yellow.
Calcium malate agar: Colonies yellowish;
aerial mycelium gray to white-gray. Soluble
pigment yellow.
Glucose-asparagine agar: Growth abun-
dant; aerial mycelium gray. Soluble yellow
pigment, especially with high nitrogen con-
centration.
nutrient Good substrate
growth, at first cream-colored, later sulfur-
vellow. Aerial mycelium white, later gray.
Glucose agar:
Potato: Growth light cream-colored, later
often yellow. Aerial mycelium white, later
slate-gray.
Gelatin: Growth yellowish-gray to gray-
ish-black. Rapid liquefaction.
Milk: Rapid coagulation and peptoniza-
tion.
Cellulose: Growth good.
Esculin: Hydrolysis.
Starch: Diastatic action strong.
Nitrate: Reduction weak.
Invertase: Negative.
Production of HS: Negative.
Temperature: Optimum 30-35°C.
Pigment: Soluble in alcohol and other or-
ganic solvents.
Antagonistic properties: Produces anti-
biotics fungichromin and actinomycin.
Habitat: Very common in soil.
Remarks: The culture described by Krain-
sky (1914) produced an aerial mycelium of a
eray color (‘like diastaticus’’). Later, how-
ever, Krainsky’s culture was found to pro-
duce a grayish-yellow aerial mycelium like
the streptomycin-producing S. griseus (Ett-
linger ef al., 1958); these authors also con-
sider the S. cellulosae strains obtained from
ATCC and NRRL as closely related to S.
griseus. Most probably, one of these cultures
was used in the studies of Tresner and Dang:
(1958), who mention a yellow-cream-buff
color of the aerial mycelium. In contrast to
these cellulosae strains now available for
comparison, Jensen (1930a) described five
soil isolates as A. cellulosae with a distine-
tive slate-gray aerial mycelium, in agree-
ment with Krainsky’s description. Krassilni-
kov (1949) considers this organism as a
variety of A. flavus.
Type culture: IMRU 3313, 3780.
49. Streptomyces chartreusis Calhoun and
Johnson, 1956 (Calhoun, Ik. M. and John-
DESCRIPTION OF SPECIES OF STREPTOMYCES
son, L. E. Antibiotics & Chemotherapy 6:
994-298, 1956).
Morphology: Aerial hyphae branch pro-
fusely producing closed or open spirals,
sinistrorse and dextrorse, depending on com-
position of medium. Spiral chains consist of
3 to 7 turns. Spores powdery, blue-gray to
blue-green, depending on the medium. Most
spirals occur singly; a few are found in
groups of two or three measuring 5 to 20 u
in length and 3 to 5 uw in width. Spores
spherical to oval, 1.0 to 1.5 uw in diameter.
Sucrose nitrate agar: Growth profuse,
raised and somewhat wrinkled, honey-col-
ored. Aerial mycelium of young colonies
white to pale gray; older colonies have blue
center.
Glucose-asparagine agar: Growth profuse,
blue-gray. Soluble pigment yellow.
Starch agar: Growth profuse. Center of
colonies blue-green, edges white. No soluble
pigment. Good hydrolysis.
Nutrient agar: Growth profuse. Center of
colonies blue-gray, edges powdery white. No
soluble pigment.
Potato: Growth light, raised with some
wrinkling. Center of colonies blue-gray with
white edges. No soluble pigment.
Gelatin: Growth blue-green in center with
white edges. Soluble pigment yellow-green
to black. Slow liquefaction. Melanin-posi-
tive.
Milk: Growth moderate, blue-gray and
white. Slow peptonization.
Nitrate broth: Blue-green ring and white
pellicle. Soluble pigment vellow-tan. Nitrate
strongly reduced.
Production of H.S: Positive.
Antagonistic properties: Produces anti-
biotic chartreusin.
Xtemarks: Ettlinger et al. (1958) considered
this species to be related to S. vzridochromo-
genes.
50. Streptomyces chibaensis Suzuki et al.,
1958 (Suzuki, 8., Nakamura, G., Okuma, K..,
195
and Tomiyama, Y.
11LA: 81-83, 1958).
Morphology: Sporophores wavy, produc-
J. Antibiotics (Japan)
ing numerous small spirals.
Sucrose nitrate agar: Growth slow, pene-
trating into medium, yellow reverse. At first
no aerial mycelium; later mycelium pro-
duced, white becoming gray, then reddish-
gray or black-buff. Soluble pigment shght
vellow.
Nutrient
cream-colored or yellow. Aerial mycelium
agar: Growth good. Reverse
powdery, white. Soluble pigment absent or
slightly yellow.
Glucose-asparagine agar: Growth good,
vellow, later becoming brown. Aerial myce-
lium powdery, white, later becoming dark
brown. Soluble pigment yellow.
Starch agar: Growth good, white, later be-
coming olive-yellow. Strong hydrolysis of
starch.
Potato: Growth good, raised. Aerial my-
celium white or cream-colored, powdery. No
change in color of plug.
Gelatin: Growth on surface and in medium
poor. No aerial mycelium. Liquefaction ab-
sent or limited. Melanin-negative.
Milk: Growth good; white to cream-col-
ored pellicle. No aerial mycelium. No soluble
pigment. Peptonization slow.
Carbon sources: Utilizes readily a variety
of carbon sources, but not inulin.
Antagonistic properties: Produces anti-
biotic cellocidin, active against gram-positive
and gram-negative bacteria; this antibiotic
also possesses anticancer properties.
Habitat: Soil in Japan.
temarks: The organism is closely related
to S. flavus.
51. Streptomyces chrysomallus Lindenbein,
1952 (Lindenbein, W. Arch. Mikrobiol. 17:
361-383, 1952).
Morphology: Substrate growth soft, con-
sisting of long, branching hyphae, with nu-
merous staining granules. Sporophores long,
194
straight; no spirals. Spores oval to elliptical;
surface smooth.
Glycerol nitrate agar: Growth light yellow.
Aerial mycelium powdery, white. Soluble
pigment golden yellow.
Glucose-asparagine agar: Growth smooth,
colorless to yellowish. Aerial mycelium
powdery, white. Soluble pigment faint yel-
low.
Glycerol malate Growth thin,
smooth, colorless to light yellow. Aerial my-
celium powdery, grayish-white.
Nutrient agar: Growth poor, shiny, golden
yellow. Aerial mycelium white, powdery.
Soluble pigment golden yellow. Melanin-
agar:
negative.
Glucose-peptone agar: Growth yellowish
with tinge of orange. Aerial mycelium gray-
ish-white. Soluble pigment lhght yellow to
golden yellow.
Starch-casein agar: Growth colorless, with
yellowish reverse. Aerial mycelium powdery,
chalk-white. Strong hydrolysis of starch. No
soluble pigment.
Potato: Growth heavy, yellow, becoming
brownish-yellow or orange. Aerial mycelium
cottony white to yellowish-white.
Gelatin: Surface growth heavy, hght to
dark yellow. Aerial mycelium white. Soluble
pigment yellow-brown to deep brown, only
in liquefied portion. Strong liquefaction.
Melanin-negative.
Milk: Growth colorless, with hight yellow
reverse. Aerial mycelium cottony, snow-
white, becoming yellowish. Coagulation
shght. Strong peptonization.
Cellulose: Growth very weak.
Production of HoS: Negative.
Antagonistic properties: Produces actino-
mycin C; some strains also produce the
antifungal cycloheximide.
Habitat: Soil.
Remarks: Ettlinger et al. (1958) considered
it as a member of the S. griseus group. A
complete description of this organism was
(1958). Frommer
also given by Frommer
THE ACTINOMYCETES, Vol. II
(1959) described a variety of this organism
under the name fumigatus. It differed from
the type species by producing a mouse-gray
aerial mycelium on synthetic media, no
aerial mycelium on potato and gelatin, and
by displaying more limited proteolytic prop-
erties.
Type culture: IMRU 3657.
52. Streptomyces cinereoruber Corbaz et al.,
1957 (Corbaz, R., Ettlinger, L., Keller-
Schierlein, W., and Zihner, H. Arch. Mikro-
biol. 25: 825-332, 1957).
Morphology: Sporophores straight; no
spirals. Spores shghtly elongated, 0.9 to 2 by
0.7 to 1 w; surface of spores smooth.
Glycerol nitrate agar: Growth thin, light
carmine-red, in 7 days dark red. Aerial my-
celium ash-gray. Soluble pigment lght car-
mine.
Glucose-asparagine agar: Substrate
erowth thin, greenish-gray to bluish-gray.
Aerial mycelium ash-gray. No soluble pig-
ment.
Gelatin: Surface growth hght carmine to
light brown. Aerial mycelium light gray. Sol-
uble pigment red-brown. Medium liquefac-
tion. Melanin-positive.
Starch agar: Vegetative growth coral-red.
Aerial mycelium ash-gray. Soluble pigment
carmine. Hydrolysis limited.
Potato: Growth lichenoid, brownish-yel-
low. Aerial mycelium ash-gray. Soluble pig-
ment bluish-gray.
Milk: Pellicle ght brown with sparse
aerial mycelium, powdery, white-gray. Coag-
ulation and peptonization. Reaction turns
acid.
Carbon utilization: Utilizes xylose, arabi-
nose, and other sugars. Does not utilize L-
rhamnose, b-fructose, raffinose, mulin, d-
sorbitol.
Antagonistic properties: Produces anti-
biotic rhodomycin.
Remarks: This organism is closely related
to S. bobiliae and S. purpurascens. S. cine-
reoruber var. fructofermentans is a variety,
DESCRIPTION OF SPECIES OF STREPTOMYCES
based on differences in sugar utilization, and
produces the antibiotic cinerubin.
53. Streptomyces cinnamomeus Benedict
et al., 1954 (Benedict, R. G., Dvonch, W.,
Shotwells OO) i... eEnidnam: ©:-Gy and Lin-
denfelser, L. A. Antibiotics & Chemotherapy
2: 591, 1952; 4: 1140, 1954).
The correct name of this organism is S.
cinnamomeus ft. cinnamomeus.
Morphology: Sporophores straight; later
descriptions indicate verticil formation.
Spores globose, 0.6 uw (Fig. 36).
Sucrose nitrate agar: Growth colorless to
white to cream-colored. Aerial mycelium
white to light cinnamon.
Glucose-asparagine agar: Growth color-
less; light greenish-yellow to dull yellowish-
orange in reverse. Aerial mycelium white to
cinnamon.
Nutrient agar: Growth cream-colored to
light lemon-yellow. No aerial mycelium. No
soluble pigment.
Oatmeal agar: Growth tough, leathery,
yellowish-green to cream-yellow. Aerial my-
celium floccose, pale violet to faint cinna-
mon. Exudate tan to white.
Starch agar: Growth colorless to brownish.
Aerial mycelium white. Hydrolysis.
Potato: Growth grayish-white to yellow-
green to light brown. Aerial mycelium light
gray to gray. No soluble pigment.
Gelatin: Growth flocculent, dirty yellow to
white. Aerial mycelium cretaceous. No solu-
ble pigment. Rapid liquefaction.
Milk: Ring light brown. Aerial mycelium
limited, white. Rapid peptonization.
Carbon utilization: Utilizes xylose, fruc-
tose, inositol, starch, dextrin, galactose, and
maltose. Does not utilize arabinose, rham-
nose, duleitol, and salicin.
Nitrate reduction: Negative.
Production of HS: Negative.
Temperature: Good growth at 25-37°C.
Antagonistic properties: Produces cinna-
mycin, a polypeptide antibiotic.
Source: Japanese soil.
195
FriGure 36. Hyphae of S. c¢nnamomeus showing
character of verticils of branches
(Reproduced from: Duggar, B. M. ef al.
N. Y. Acad. Sci. 60: 85, 1954).
sporogenous
Ann.
Remarks: Pridham ef al. (1956) described
a second form under the name of S. cinna-
momeus f. azacoluta; it produced a shell-pink
aerial mycelium on starch agar and an anti-
biotic, duramycin.
Type culture: IMRU 3664.
54. Streptomyces cinnamonensis Okami,
1953 (Okami, Y., Maeda, K., Kosaka, H..,
Taya, O., and Umezawa, H. Japan. J. Med.
Se. Biol. 6: 87-90, 1953).
Morphology: Sporophores long, flexible,
hooked, but no true spirals. Spores elliptical
to oval.
Nutrient agar: Growth colorless to dark.
No aerial mycelium. Soluble pigment absent
or slightly brown.
Glycerol agar: Growth colorless. Scant
white aerial mycelium or white with pale
cinnamon-pinkish to light brownish-vina-
ceous tinge. No soluble pigment.
Glucose-asparagine agar: Growth colorless
to light Aerial
white to white-pinkish-cinnamon. No soluble
cream-colored. mycelium
pigment.
196
Potato: Growth dark to light cream-col-
ored. No aerial mycelium. No soluble pig-
ment; later, black pigment produced around
growth.
Gelatin: Growth colorless to dark brown-
ish. Aerial mycelium in white
patches. Soluble pigment brown. No or very
form of
slow liquefaction.
Milk: Growth cream-colored to brownish
surface ring. Aerial mycelium absent or
scant white. Soluble pigment
slightly brown. Coagulation and peptoniza-
tion absent or very slow.
Starch agar: Growth colorless. Aerial my-
absent or
celium white with pinkish tinge. No soluble
pigment. Hydrolysis good.
Cellulose: No growth.
Nitrate reduction: None.
Production of H.S: Positive.
Carbon utilization: Sucrose, mannose, dex-
trin, galactose, glycerol, fructose, glucose,
maltose, mannitol, xylose, and sodium suc-
cinate utilized. Arabinose, esculin, rhamnose,
dulcitol, sodium acetate, mulin, lactose, sali-
cin, and raffinose not utilized.
Antagonistic properties: Produces an anti-
biotic active against mycobacteria and iden-
tical with actithiazice acid or thiozolidone.
temarks: The culture resembles S. roseo-
chromogenes in color of growth and in no or
slow liquefaction of gelatin. It differs in the
lack of spiral formation and of nitrate re-
duction. Gause et al. (1957) described a
variety of this organism under the name of
A. cinnamonensis var. proteolyticus. A. dag-
and A. fumanus described by
these authors apparently also belong to this
hestanicus
group, although they differ from it in some
respects. According to Benedict and Prid-
ham (1959) a group of cooperators consid-
ered this organism as S. cinnamonensis, S.
pirginiae, S. acidomyceticus, S. roseochromo-
genes, and S. lavendulae; an opinion was ex-
pressed that all of these are probably re-
lated to S. lavendulae.
Type culture: ATCC 12,308.
THE ACTINOMYCETES, Vol. II
59. Streptomyces circulatus (krassilnikov,
1941) Waksman (Krassilnikov, N. A. Actino-
mycetales. Izvest. Akad. Nauk. SSSR, Mos-
kau, p. 60, 1941).
Morphology: Sporophores produce verti-
cils with spiral-shaped short branches. Spores
cylindrical or oblong, 1.5 by 0.7 wu, some
rounding up with age of culture.
Synthetic agar: Growth good, colorless.
Aerial mycelium abundant, white.
Nutrient Growth weak. No aerial
mycelium.
Gelatin: Liquefaction weak.
Milk: No coagulation; slow peptonization.
Starch: Hydrolysis weak.
Cellulose: No growth.
Paraffin: Growth good. Aerial mycelium
agar:
white.
Nitrate reduction: Weak.
Sucrose: No inversion.
Antagonistic properties: Limited.
Habitat: Soil.
56. Streptomyces citreus (Xrainsky, 1914)
Waksman and Henrici, 1948 (Krainsky, A.
Centr. Bakteriol. Parasitenk. Abt. IL, 41:
684, 1914; Waksman, 8S. A. and Curtis, R.
E. Soil Sci. 1: 116, 1916; 8: 121, 1919). Not
Actinomyces citreus Gasperini, 1894.
Morphology: Sporophores form long nar-
row, open spirals, dextrorse. Spores spheri-
cal to oval, 1.2 to 1.5 by 1.2 to 1.8 yp.
Sucrose nitrate agar: Growth abundant,
raised, wrinkled, citron-yellow. Aerial myce-
lium white to citron-yellow. No soluble pig-
ment.
Malate-glycerol agar:
vellow. Aerial mycelium white with mouse-
gray tinge. No soluble pigment.
Glucose-asparagine agar: Growth glossy,
olive-yellow; center elevated. Aerial myce-
lium white to pinkish. No soluble pigment.
Nutrient agar: Growth restricted, green.
No aerial mycelium. No soluble pigment.
Potato: Growth yellowish to gray. Aerial
mycelium white. No soluble pigment.
Gelatin: Surface growth restricted, yellow-
Growth creamy to
DESCRIPTION OF SPECIES OF STREPTOMYCES
ish. Aerial mycelium white. Liquefaction me-
dium. Melanin-negative.
Milk: Surface growth cream-colored. Co-
agulation followed by rapid peptonization.
Starch media: Growth abundant, citron-
yellow to yellowish-green. Aerial mycelium
pinkish. Rapid hydrolysis of starch.
Cellulose: No growth.
Invertase: Positive.
Nitrate: Sight reduction to nitrite.
Production of HS: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Negative.
Habitat: Garden soil.
Remarks: Since Krainsky’s culture was
not available for comparison, the above de-
scription is based upon that of Waksman and
Curtis (1916) and Waksman (1919); some dif-
ferences exist between this description and
that of Krainsky. Ettlinger et al. (1958) con-
sidered this culture as a strain of S. griseus.
KrassilInikov (1949) considers this organism
as similar to Gasperini’s culture, both being
looked upon as varieties of A. flavus.
Type culture: IMRU 3574.
57. Streptomyces clavifer (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores long, straight,
some terminating in club-shaped structures.
Spores cylindrical, 1.5 by 1.0 yp.
Sucrose nitrate agar: Growth gray to
brick-red. Aerial mycelium white, sprinkled
with cinnamon-drab. Soluble pigment. yel-
lowish to brown.
Potato: Growth wrinkled, gray to orange
to brown. Aerial mycelium gray to olive-
buff. Color of plug gray to brown.
Gelatin: Growth gray to buff. Aerial my-
celium white. Medium liquefaction. Soluble
pigment yellow to reddish-yellow.
Starch: Hydrolysis.
Tyrosinase reaction: Positive.
Nitrate: No reduction.
Temperature: Fair growth at 37.5°C.
197
Habitat: Limed soil and common scab of
potatoes.
temarks: Kutzner (1956) described the
original culture obtained from CBS as pro-
ducing an ash-gray aerial mycelium without
spirals, and as melanin-negative; this culture
was indistinguishable from S. craterifer also
CBS. Krassilnikov (1949)
considered it as a variety of A. scabies.
obtained from
58. Streptomyces coelicolor (Miiller, 1908)
Waksman and Henrici emend. Kutzner and
Waksman (Miller, R. Centr. Bakteriol.
Parasitenk. Abt. I, Orig. 46: 195, 1908;
Kutzner, H. J. and Waksman, S. A.
J. Bacteriol. 78: 528-538, 1959).
Synonyms:
Streptothrix coelicolor Miller (Miller,
1908).
Actinomyces albidoflavus (strain Hohle,
CBS).
Actinomyces alni (strain v. Plotho, CBS).
Streptomyces canescus Hickey et al.
(Hickey et al., 1952, NRRL 2419).
Possible synonyms:
Gause et al. (Gause et al., 1957).
KrassiInikov (Krassilnikov,
A. cyaneofuscatus
A. levoris
1958).
Not S. violaceoruber.
Morphology: Sporophores of most strains
short, arranged in small tufts, wavy; no
spirals. Spores spherical to ellipsoidal; sur-
face smooth.
Agar media: Substrate growth on most
media colorless or atypical yellowish-brown-
ish; sometimes pinkish-red, especially in the
lower part of the slants. Aerial mycelium
colored grayish-yellow, often with a green-
ish or pinkish shade. Soluble pigment on
most media either absent or yellowish-
brown. Blue pigment is produced by some
strains on glucose-caletum malate-NHyNOs
agar, mannitol-calcium malate-peptone agar,
or glucose-peptone agar.
Potato: Growth abundant, lichenoid. Aer-
to yellow.
ial mycelium powdery, white
Characteristic formation of greenish-blue to
198
sky-blue soluble pigment by several strains;
it may later become deep blue or blue-violet.
Addition of glycerol delays pigment forma-
tion.
Gelatin: Good growth. Rapid liquefaction.
No soluble pigment.
Milk: No coagulation, rapid peptoniza-
tion, complete within 15 days at 22-27°C;
coagulation within 3 to 5 days, followed by
peptonization at 36°C.
Starch hydrolysis: Strong.
Nitrate reduction: Positive; none reported
for S. canescus by Hickey et al. (1952).
Carbon sources: Utilizes L-xylose, L-arabi-
nose, p-fructose, D-galactose, pb-mannitol,
salicin; does not utilize L-rhamnose or raffi-
nose; most strains do not utilize sucrose.
Hemolysis of blood: Rapid at 37°C.
Production of H.S: Negative.
Antagonistic properties: Active upon sev-
eral fungi and yeasts; all strains as far as
tested produce polyene antibiotics. S. griseus
(Krainsky) Waksman and Curtis (1916)
probably belongs to this species, since it 1s
now known to produce an antifungal agent
of the polyene type.
Ecology: S. coelicolor is widely distributed
in nature. In a search for polyene-producing
organisms, Pledger and Lechevalier (1955—
1956) found 26 strains among 93. isolates
which produced polyenes and which can be
regarded as belonging to this species. Among
the 382 subgroups of Kutzner (1956), the
S. coelicolor subgroup was the one which
comprised most Heymer (1957)
found this organism strikingly often on the
skin and in the tonsils of men. The first
culture of this species isolated by Miller
(1908) and the ascosin-producing organism
GS. canescus) were found as chance con-
strains.
taminants; this indicates the wide distribu-
tion of the organism in air. The relationship
of blue pigment-forming bacteria, designated
as Actinobacillus and Actinococcus, to this
was Beijerinck
organism discussed — by
(1913a).
THE ACTINOMYCETES, Vol. II
Numerous cultures isolated by different
investigators and described as S. coelicolor
belong to S. violaceoruber. Others, however,
such as A. tricolor Wollenweber, are related
to S. coelicolor.
Type culture: A strain of this organism
was deposited by R. Miiller in the CBS.
59. Streptomyces collinus Lindenbein, 1952
Lindenbein, W. Arch. Mikrobiol. 17: 361—
383, 1952).
Morphology:
Spores oval.
Glycerol nitrate agar: Growth yellow-
brown to red-brown. Aerial mycelium chalk-
white. Soluble pigment yellow-brown, later
becoming reddish-brown.
Glucose-asparagine agar: Growth yellow-
brown to purple-red. Aerial mycelium chalk-
white, later ash-gray. Soluble pigment car-
mine-red, later brown-red.
Sporophores form. spirals.
Glycerol malate agar: Growth yellow-
brown to red-brown. Aerial mycelium vel-
vety, chalk-white. Soluble pigment yellow-
brown.
Nutrient agar: Growth dark brown. Aerial
mycelium powdery, gray-white. Soluble pig-
ment dark brown. Melanin-positive.
Glucose-peptone agar: Growth yellow-
brown and red. Aerial mycelium velvety,
white. Soluble pigment chestnut-brown.
Starch media: Growth reddish to orange.
Aerial mycelium white. Hydrolysis medium.
Potato: Growth good. Aerial mycelium
white. No soluble pigment.
Gelatin: Growth dark brown. No aerial
mycelium. Soluble pigment dark brown.
Liquefaction rapid.
Milk: Growth good; dark brown reverse.
Aerial mycelium white, later ash-gray. Solu-
ble pigment dark brown. No peptonization.
Cellulose: Growth good, colorless.
Antagonistic properties: Produces an anti-
biotically active pigment.
Habitat: Soil.
Remarks: Closely related to S. erythro-
chromogenes. Gause et al. (1957) described a
DESCRIPTION OF SPECIES OF STREPTOMYCES
similar form under the name of A. albovina-
ceus.
60. Streptomyces coroniformis (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight, some
long and some short. Spores oval, 0.8 by
0.6 p.
Sucrose nitrate agar: Growth in form of
discrete colonies partially coalescing, gray to
greenish. Aerial mycelium white, covering
edges of growth.
Nutrient potato agar: Growth wrinkled,
grayish. No aerial mycelium.
Potato: Growth raised, grayish. Aerial
mycelium white. Plug pigmented brownish
around and under growth.
Gelatin: Growth fair. Liquefaction slow if
any.
Milk: A few colonies on surface. No coagu-
lation; peptonization limited.
Starch: No hydrolysis or trace.
Nitrate: Limited reduction to nitrite.
Tyrosinase reaction: Negative.
Temperature: Growth fair at 37.5°C
Habitat: Potato scab.
61. Streptomyces craterifer (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Straight sporophores form
terminal branches, dichotomously forked.
Spores rectangular, 1.8 to 0.9 by 1.0 to 0.8 wu.
Sucrose nitrate agar: Growth lichenoid,
abundant, colorless; aerial mycelium mouse-
gray. Numerous guttation drops, which
leave blackish craters on surface of growth.
Nutrient agar: Growth colorless; aerial
mycelium scant, white. No soluble pigment.
Starch agar: Growth spreading, thin, col-
orless; no aerial mycelium. Starch hydro-
lyzed.
Potato: Growth cream-colored; aerial my-
celium white to mouse-gray. Color of plug
unchanged.
Gelatin: Surface growth wrinkled; aerial
199
mycelium white. No
soluble pigment.
Milk: Surface growth cream-colored. No
coagulation, rapid peptonization.
Rapid liquefaction.
Tyrosinase reaction: Negative.
Nitrate reduction: Positive.
Temperature: Only
See
Habitat: Raised, smooth scab.
Type culture: IMRU 3373.
62. Streptomyces cyaneus (Krassilnikoy,
1941) Waksman (Krassilnikov, N. A. Actin-
omycetales. Izvest. Akad. Nauk. SSSR, Mos-
kau, p. 14, 1941).
Morphology: Sporophores produce open
spirals (sinistrorse), with 2 to 3
shght growth at
turns in
ach. Spores oval, seldom spherical, 0.6 to
0.8 by 0.6 uw.
Agar media: Growth pigmented blue at
both acid and alkaline reactions. The pig-
ment does not dissolve into medium. Aerial
mycelium well developed, downy, bluish-
gray to blue-green in color.
nitrate Colonies at first
smooth, becoming lumpy, leathery, com-
pact, and covered with well developed blue-
gray aerial mycelium.
Gelatin: Strong
positive.
Milk: Peptonization without prior coagu-
Sucrose
agar:
liquefaction. Melanin-
lation.
Starch: Hydrolysis weak.
Cellulose: No growth.
Nitrate reduction: Negative.
Sucrose inversion: Negative.
Antagonistic properties: Weak.
Type culture: IMRU 3761.
63. Streptomyces cyanoflavus Funaki and
Tsuchiya, 1958 (Funaki, M., Tsuchiya, F.,
Maeda, K., and Kamiya, T. J. Antibiotics
(Japan) LLA: 143-149, 1958).
Morphology: Sporophores straight, form-
ing many branches, but no spirals.
Sucrose nitrate agar: Growth colorless to
pale yellowish-brown. Aerial mycelium white
to light greenish-gray. No soluble pigment.
200 THE ACTINOMYCETES, Vol. II
Glucose-asparagine agar: Growth pale
green to yellowish-brown. Aerial mycelium
brownish-white to brownish-gray. Soluble
pigment hght blue to yellowish-brown.
Calcium malate agar: Growth light brown-
ish to brown. Aerial mycelium white to gray.
Soluble pigment hight
brown.
Nutrient agar: Growth yellowish-brown to
brown. Aerial mycelium grayish-white. No
soluble pigment.
Yeast extract agar: Growth pale yellow to
brown. Aerial mycelium light olive-gray.
Soluble pigment brown.
Potato: Growth yellow to brown. Aerial
greenish-blue — to
mycelium brownish-gray. Soluble pigment
dark brown.
Gelatin: Growth yellow. Soluble pigment
yellow to brownish-yellow. Gelatin lique-
fied.
Milk: Produces a sedimented growth with-
out any soluble pigment. Milk coagulated
but not peptonized.
Carbon utilization: Utilizes various sugars
and salts of organic acids, but not xylose,
acetate, or citrate.
Antagonistic properties: Produces blue
antibiotic ¢cyanomycin, active against gram-
positive and gram-negative bacteria; also
produces aureothricin-like substances.
64. Streptomyces cylindrosporus (kXrassil-
nikov, 1941) Waksman (Krassilnikoy, N. A.
Actinomycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 57, 1941).
Morphology: Sporophores straight,
branched. Spores cylindrical or oblong, 1.0
bowler. oy O57 jes
Sucrose nitrate agar: Colonies velvety,
dark brown or chocolate. Aerial mycelium
white-gray to brown-gray. Soluble pigment
brown.
Nutrient agar: Growth dark brown. Aerial
mycelium white. Soluble pigment brown.
Glucose-asparagine agar: Growth brown.
Aerial mycelium white-gray. No soluble pig-
ment.
Gelatin: Weak liquefaction. Melanin-posi-
tive.
Milk: Coagulation limited, peptonization
weak; color of milk brown to almost black.
Potato: Substrate growth brown. Aerial
mycelium light gray. Soluble pigment brown.
Starch: Weak hydrolysis.
Cellulose: Limited, colorless growth. Aer-
ial mycelium white.
Nitrate reduction: Positive.
Sucrose inversion: Negative.
Antagonistic properties: None.
Habitat: Soil.
temarks: The description of the organism
has been supplemented by Hoffmann (1958).
It appears to be related to S. vinaceus, S.
purpureochromogenes, and S. purpeofuscus.
Gause et al. (1957) described a related form
as A. wmbrinus.
Type culture: IMRU 3764.
65. Streptomyces diastaticus (Krainsky)
Waksman and Henrici (Krainsky, A. Centr.
Bakteriol. Parasitenk. Abt. II, 41: 682,
1914).
Morphology: Sporophores form tight spi-
rals. Spores oval or spherical, 1.0 to 1.2 by
LAstow5iueChies: 3738):
Sucrose nitrate agar: Growth thin, gray.
Aerial mycelium white, becoming drab gray.
Calcium malate agar: Colonies 2 to 4 mm,
yellowish when old. Aerial mycelium gray,
with white outer zone; white specks fre-
quently produced in gray mycelium.
Glucose-asparagine agar: Growth yellow-
ish, spreading. No aerial mycelium.
Growth
Aerial mycelium white, then gray. Soluble
Nutrient agar: cream-colored.
brown pigment.
Potato: Growth white-gray. Aerial myce-
lium gray and white.
Gelatin: Liquefaction, with small, cream-
colored flakes in liquefied part.
Milk: Brownish ring. Coagulation and
slow peptonization.
Starch thin, colorless,
agar: Growth
—_—__
DESCRIPTION OF
SPECIES OF STREPTOMYCES 201
FrGuReE 37. Sporophores of S. diastaticus, X 4,500, showing uniform density over whole surface (Cour
tesy of E. Baldacci, University of Milan, Italy).
spreading. Aerial mycelium gray. Ready hy-
drolysis.
Cellulose: Good growth.
Invertase: Negative.
Nitrate reduction: Weak.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Limited.
Habitat: Soil.
temarks: This species was redescribed by
Duché (1934) under the name A.
ticus. Baldacci et al. (1955) raised this spe-
roseodiasta-
cies to the status of a ‘“‘series.’? Several new
species or varieties were created: A. virido-
diastaticus, A. diastaticus var. ardesicicus, A.
diastaticus var. venezuelae, A. rubrocyano-
diastaticus var. impiger and var. piger.
Type culture: IMRU 3315.
66. Streptomyces diastatochromogenes
(Krainsky, 1914; Waksman and Curtis,
1916) Waksman and Henrici, 1948 (krain-
sky, A. Centr. Bakteriol. Parasitenk. Abt.
IT., 41: 683, 1914).
Morphology: According to Waksman and
Curtis (1916), sporophores are straight. Ac-
cording to Jensen (1930), sinistrorse spirals
are produced. Spores spherical or oval, 1.2 yu.
Growth
Aerial
later ash-gray. Soluble
Sucrose nitrate colorless,
agar:
later yellowish-brown. mycelium
abundant, white,
pigment yellowish to light brown.
Sucrose malate agar: According to IKrain-
sky, growth colorless, with gray aerial myce-
lium. When glucose is added, center of aerial
mycelium is colored yellowish, with gray
margin.
Glucose-asparagine agar: Growth color-
less, with gray aerial mycelium.
Nutrient agar: Aerial mycelium white to
gray. Soluble pigment brownish to coffee-
brown. Melanin-positive.
Potato: Growth light gray, later grayish-
black. Aerial mycelium white to gray. Solu-
ble pigment black.
Gelatin: Growth cream-colored to vellow-
ish-brown. Aerial mycelium scant white. Sol-
202
THE ACTINOMYCETES, Vol. II
FIGURE 38. Sporophores of S. diastaticus, * 15,000 (Courtesy of E. Baldacci, University of Milan,
Italy).
uble pigment brown. Liquefaction fairly
Habitat: Very common in soil.
rapid.
Remarks: Krassilnikov (1949) considered
this species as a variety of A. chromogenes.
Type culture: ATCC 12,309.
Starch: Hydrolysis weak.
Cellulose: No growth.
Nitrate: Reduction to nitrite strong. 67. Streptomyces echinatus Corbaz et al.,
1957 (Corbaz, R., Ettlinger, L., G&asmann,
Temperature: Optimum 35°C. E., Ixeller-Schierlein, W., Kradolfer, F.,
Antagonistic properties: Very strong. Neipp, L., Prelog, V., Reusser, P., and
Tyrosinase reaction: Positive.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Zahner, H. Helv. Chim. Acta 40: 199-204,
1957).
Morphology: Sporophores produce verti-
cils on sterile aerial hyphae, with open, ir-
regular spirals. Spores elliptical to oval; sur-
face of spores covered with long, thin spines
(Pies).
Glycerol nitrate agar: Growth greenish-
yellow to citron-yellow to light green. Aerial
mycelium white, changing to yellow, to ash-
gray. Soluble pigment greenish-yellow to
erass-green.
Glycerol malate agar: Growth pale yellow,
turning greenish-yellow. Aerial mycelium
white to pale yellow. No soluble pigment.
Glucose-asparagine agar: Growth golden
vellow to greenish-yellow to greenish-gray.
Aerial mycelium ash-gray to reddish-violet.
No soluble pigment.
Nutrient agar: Growth light yellow. No
aerial mycelium. No soluble pigment.
Glucose-peptone agar: Growth
Aerial mycelium white in center, brownish
yellow.
on periphery, changing to ash-gray. Soluble
pigment golden yellow.
Starch agar: Growth yellow. Aerial myce-
hum ash-gray. No hydrolysis, or at most,
traces.
Gelatin: Substrate growth dark brown.
Aerial mycelium greenish-gray. Soluble pig-
ment dark brown. No liquefaction after 31
days. Melanin-positive.
Potato: Growth greenish to raven-black.
Aerial mycelium limited, white-gray to
bluish-gray. Soluble pigment brownish to
pitch-black.
Milk: Good coagulation and peptoniza-
tion.
Tyrosinase reaction: Positive.
Carbon utilization: Xylose, lactose, raffi-
nose, acetate, and succinate—positive. Su-
crose, inulin, duleitol, salicin, and sodium
citrate—negative.
Temperature: Develops well at 18-40°C.
Antagonistic properties: Produces echino-
mycin.
205
temarks: Closely related to S. griseoflavus
and S. flaveolus.
68. Streptomyces elasticus (Séhngen and
Fol, 1914) Waksman (Séhngen, N. L. and
Fol, J. G. Centr. Bakteriol. Parasitenk. Abt.
IT, 40: 87-98, 1914).
Morphology: Mycelium typical, branched.
Short, motile rods observed in young cul-
tures. Spores white, round, diameter about
1 uw, double that of the mycelium.
Agar Growth yellowish-white.
Aerial mycelium snow-white.
Gelatin: Growth yellow-brown.
media:
Carbon utilization: Glucose, glycerol, ethyl]
alcohol, mannitol, organic acids,
salts readily assimilated.
calelum
Sucrose inversion: Positive.
Urea: Produces urease.
Paraffin: Utilized.
tubber: Utilized readily.
Temperature: Optimum 28°C, maximum
33°C, destroyed at 65°C in 5 minutes.
Habitat: Soil.
69. Streptomyces endus Gottlieb and Car-
ter, 1956 (Gottlieb, D. and Carter, H. E.
U.S. Patent 2, 746, 902, May 22, 1956).
Morphology: Sporophores formed along
entire length of mycelium, at right angles
to it. Compact spirals produced, often with
10 loops. Young hyphae 0.7 to 1.0 uw in
diameter; old hyphae 1.25 to 1.50 and even
2.0) us.
Sucrose nitrate agar: Substrate growth
has color of medium, later turning dark.
Aerial mycelium white, changing to light
gray, then to dark gray. No soluble pigment.
Gelatin: Slow and only slight liquefaction.
No soluble pigment. Melanin-negative.
Starch: Hydrolysis rapid.
Potato: Growth good. Aerial mycelium
hight gray. No soluble pigment.
Milk: Coagulation; no visible peptoniza-
tion.
Carbon utilization: Utilizes starch, man-
nose, dextrin, glucose, arabinose, maltose,
204
and levulose. Poor growth with galactose,
lactose, citric acid, malic acid, succinic acid,
and Does utilize
sorbitol, duleitol, inositol, or paraffin.
cellulose. not sucrose,
Antagonistic properties: Produces an
antibiotic, endomycin, active largely upon
fung.
Remarks: Tresner and Backus (1956)
consider this organism as a variant of the
S. hygroscopicus group.
70. Streptomyces erythraeus (Waksman
and Curtis, 1916) Waksman and Henrici,
1948 (Waksman, 8S. A. and Curtis, R. E.
Soil Sci. 1: 99, 1916; Waksman, S. A. Soil
sci: 8: 112, 1919).
Morphology: Fine, monopodially
branched aerial mycelium; numerous sporo-
phores with open and closed spirals. Spores
spherical to oval, 0.7 to 0.8 uh, smooth (PI.
Il im).
Sucrose nitrate agar: Growth yellowish,
later becoming red. Pigment insoluble in
medium. Aerial mycelium thick, white to
pale rose.
Glucose-asparagine agar: Growth abun-
dant, spreading, cream-colored, later turn-
ing brown chiefly on surface; center raised,
lobate margin.
Nutrient agar: Substrate growth cream-
colored. No soluble pigment.
Potato: Growth wrinkled, cream-colored,
becoming yellowish to red to purplish.
Melanin-negative.
Gelatin: Growth abundant, dense, gray
with pinkish tinge, chiefly on surface of
slowly liquefied portion. No soluble pigment.
Milk: Surface zone yellowish. Limited
coagulation and positive peptonization.
Starch media: Growth cream-colored with
faint greenish tinge. Hydrolysis.
Cellulose: Growth brick-red.
Invertase: None.
Nitrate: Reduction to nitrite only with
starch.
Production of HoS: Negative.
Temperature: Optimum 25°C.
THE ACTINOMYCETES, Vol. II
Antagonistic properties: Marked. Pro-
duces erythromycin A and B.
Habitat: Soil.
Remarks: According to Ettlinger et al.
(1958) S. rimosus and S. roseochromogenes
belong to this group. Krassilnikov (1949)
considers this organism as a variety of A.
ruber. A closely related, melanin-positive
culture has been described as a new species,
S. bottropensis (Brit. Pat. 762, 736, Nov. 19,
1953).
Type culture: IMRU 3737; ATCC 11,635.
71. Streptomyces erythrochromogenes
(Krainsky, 1914) Waksman and Henrici,
1948 (kxrainsky, A. Centr. Bakteriol. Para-
sitenk. Abt. II, 41: 679-682, 1914).
Different strains of this organism have
been studied by Krainsky (1914), Waksman
and Curtis (1916), Jensen (1930), and Okami
and Suzuki (1958).
Morphology : Sporophores flexible, curved;
spiral formation abundant according to
Jensen and Okami and Suzuki. Waksman
and Curtis reported no spirals. Spores oval.
nitrate agar: Growth at first
cream-colored, later turning red to violet to
purple. Aerial mycelium white to light gray.
Sucrose
Soluble pigment red to red-violet according
to Jensen.
Calcium agar: Growth red to
violet. Aerial mycelium grayish, with white
margin.
malate
Glucose-asparagine agar: Aerial mycelium
eray to white. Soluble pigment red.
Nutrient agar: Growth yellowish-gray to
light brown. Aerial mycelium white to light
gray. Soluble pigment brown to deep brown.
Starch agar: Aerial mycelium gray. Solu-
ble pigment rose-colored. Diastatic action
weak.
Potato: Growth yellowish-gray, later al-
most black. Aerial mycelium gray. Soluble
pigment black. Melanin-positive.
Gelatin: Growth yellowish to light purple.
Liquefaction very slow. Soluble pigment
brown.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Cellulose: Growth slow or none.
Invertase: Negative.
Nitrate: Reduction slight.
Pigment: Soluble in water, not in organic
solvents.
Temperature: Optimum 380°C.
Antagonistic properties: Produces sarko-
mycin (Okami and Suzuki).
Habitat: Not very common in soil.
72. Streptomyces eurocidicus Okami et al.,
1954 (Okami, Y., Utahara, R., Nakamura,
S.,and Umezawa, H. J. Antibioties (Japan)
TA: 101-102, 1954).
Morphology: Aerial mycelium — short,
branched. Sporophores straight, without
spirals; sometimes atypical verticils are
produced.
Glycerol nitrate agar: Growth colorless
to yellowish-brown. Aerial mycelium scant,
thin, white. No soluble pigment.
Glucose-asparagine agar: Growth colorless
to vellowish-brown. Aerial mycelium white
with yellowish tinge. Soluble pigment absent
or slightly brown.
Nutrient agar: Growth vellowish-brown
to black. Soluble pigment brown. Melanin-
positive (?).
Starch: Good hydrolysis.
Potato: Growth wrinkled, brownish-yel-
low. Aerial mycelium absent or thin white.
No soluble pigment.
Gelatin: Growth yellowish-brown. Soluble
pigment brown. No liquefaction.
Milk: Surface ring vellowish-brown.
Nitrate: No reduction.
Tyrosinase: Doubtful.
Antagonistic properties: Produces an
antifungal substance, eurocidin, and anti-
bacterial substances tertiomycin and azomy-
cin.
73. Streptomyces eurythermus Corbaz et al.,
1955
K., Keller-Schierlein, W., Neipp, L., Prelog,
V., Reusser, P., and Zihner, H. Helv. Chim.
Acta 38: 1202-1209, 1955).
(Corbaz, R., Ettlinger, L., Gaiumann,
205
Morphology: Substrate growth consists
of long hyphae. Aerial mycelium abundant,
gray. Sporophores broom-shaped. Spores
egg-shaped to spherical, smooth, 0.8 to 1.0
by 0.6 to 0.7 u.
Glycerol nitrate agar: Growth postulate
light brown. Aerial mycelium sparse, white-
gray, changing to ash-gray. Soluble pig-
ment brown.
Nutrient agar: Growth brownish-yellow.
Aerial mycelium ash-gray. Soluble pigment
reddish-brown.
Glucose-asparagine agar: Growth thin,
whitish yellow. Aerial mycelium white-gray
to ash-gray. Soluble pigment chestnut-
brown.
Starch agar: Growth golden yellow. Aerial
mycelium velvety, at first snow-white, later
gray. Soluble pigment light brown. Rapid
hydrolysis.
Gelatin: Growth sparse. Soluble pigment
dark brown. Rapid liquefaction.
Potato: Growth — lichenoid,
yellow. Aerial mycelium milky-white, be-
brownish-
coming ash-gray. Soluble pigment brownish
to pitch-black.
Milk: Brown surface ring. Aerial mycelium
ash-gray. Soluble dark
Coagulation and peptonization positive.
pigment brown.
Carbon utilization: Xylose, arabinose,
fructose, galactose, saccharose, maltose,
lactose, mannitol, salicin well utilized.
Rhamnose, inulin, sorbitol, dulcitol, meso-
inositol not utilized. Some strains use acetate,
citrate, and succinate.
Temperature: Poor growth at 18°C; very
good growth at 30°C; good growth but no
aerial mycelium at 58°C.
Antagonistic properties: Produces a basic
antibiotic, angolamycin, related to erythro-
mycin.
Habitat: Soil.
Closely anti-
temarks: related to S.
bioticus.
74. Streptomyces exfoliatus (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
206
(Waksman, 8. A. and Curtis, R. E. Soil Sei.
PetG6; AGI6 S82 1211919).
Morphology: Colony has tendency to
crack and surface growth to exfohate and
peel off. Sporophores usually straight or
slightly wavy; on some media there is a
tendency to produce spirals. Spores oval, 1.0
tomleorbyalk2sto-lcSin-
Sucrose nitrate agar: Growth smooth,
colorless, becoming brown to blue. Aerial
mycelium white.
Malate-glycerol agar: Growth cream-
colored. Aerial mycelium white. No soluble
pigment.
Glucose-asparagine agar: Growth cream-
colored, turning brown. Aerial mycelium
white, appearing late.
Nutrient agar: Growth No
aerial mycelium. Soluble pigment absent or
colorless.
brownish.
Potato: Growth wrinkled, gray, becoming
brown. No aerial mycelium. No soluble pig-
ment.
Gelatin: Growth cream-colored. Aerial
mycelium white or absent. Liquefaction
faint to fair. Melanin-negative.
Milk: Cream-colored ring. Soft coagula-
tion and slow peptonization.
Starch media: Growth restricted, gray
becoming brown. Aerial mycelium light
buff-gray. Hydrolysis of starch medium,
incomplete.
Invertase: Positive.
Cellulose: Growth good.
Nitrate reduction: Positive.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Positive.
Habitat: Soil.
femarks: Krassilnikov (1949) considered
this organism as a variety of one of the
chromogenic groups.
Type culture: IMRU 3316.
75. Streptomyces felleus Lindenbein, 1952
(Lindenbein, W. Arch. Mikrobiol. 17: 361-
383, 1952).
THE ACTINOMYCETES, Vol. II
Morphology: Sporophores long, straight,
branching. Spores spherical, smooth.
Glycerol nitrate agar: Growth smooth,
o
yellow-brown. Aerial mycelium velvety
eray-white. Soluble pigment —yellowish-
brown.
Glucose-asparagine agar: Growth colorless
to brownish-yellow. Aerial mycelium gray-
white. Soluble pigment brownish.
Glycerol malate agar: Growth
to yellowish. Aerial mycelium powdery,
eray-white. Soluble pigment
brown.
Nutrient
ish-yellow
Soluble pigment
Melanin-negative.
Glucose-peptone agar: Growth yellowish-
brown. Aerial mycelium gray-white. Soluble
pigment light brown.
Starch media: Growth lichenoid, colorless.
Aerial mycelium white. No soluble pigment.
Hydrolysis strong.
colorless
yellowish-
agar: Growth colorless, brown-
reverse. No aerial mycelium.
light brownish-yellow.
Potato: Growth brownish-yellow. No
aerial mycelium. Soluble pigment absent or
pinkish.
Gelatin: Growth colorless. No aerial myce-
lium. No soluble pigment. No lquefaction
by one strain, positive by another.
Milk: Growth brownish to orange. Aerial
mycelium gray-white. Peptonization me-
dium.
Cellulose: No or weak growth.
Production of HS: Negative.
Odor: Typical earthy.
Taste: Gall-bitter.
Antagonistic properties: Produces anti-
biotic picromycin.
Remarks: Related to S. fimicarius. Ett-
linger et al. (1958) considered this organism
as belonging to S. olivaceus.
Type culture: IMRU 3659.
76. Streptomyces fervens DeBoer et al.,
1959 (DeBoer, C., Dietz, A., Evans, J. 8.,
and Michaels, R. M. Antibiotics Ann. 1959-
1960, pp. 220-226).
DESCRIPTION OF SPECIES OF STREPTOMYCES
Morphology: Sporophores monoverticil-
late or biverticillate. Pigment granules
present in mycelium.
Sucrose nitrate agar: Growth faint-pink.
Aerial mycelium pink. No soluble pigment.
Calcium malate agar: Growth © pink.
Aerial mycelium trace, pink. No soluble pig-
ment.
Glucose-asparagine
Aerial mycelium pink. Soluble pigment pale
agar: Growth pink.
yellow.
Starch nutrient agar: Growth red-pink.
Aerial mycelium pink. Soluble pigment tan.
Casein digest-beef extract agar: Growth
red. Aerial mycelium pink.
Gelatin: Liquefaction medium. Soluble
pigment brown.
Milk: Growth cream-pink.
Aerial mycelium trace pink. No coagulation.
No peptonization.
Production of H.S: Positive.
Starch: Hydrolyzed.
Carbon utilization: Utilizes various carbo-
hydrates, glycerol, inositol, starch, certain
brown to
organic acids (acetate, citrate, succinate) ;
does not utilize d-xylose, rhamnose, lactose,
l-arabinose, formic, oxalic, and tartaric acids.
Antagonistic properties: Produces anti-
biotic fervenulin, active against various
microbes and tumors.
Habitat: Soil in California.
77. Streptomyces filamentosus Okami et al.,
1953 (Okami, Y., Okuda, T., Takeuchi, T..,
Nitta, K., and Umezawa, H. J. Antibiotics
(Japan) 6A: 153-157, 1953).
Morphology: Sporophores straight, long,
without spirals. Spores oval to elliptical.
Sucrose nitrate agar: Growth colorless.
Aerial mycelium abundant, cottony, white.
No soluble pigment.
Glucose-asparagine agar: Growth color-
less. Aerial mycelium abundant, white with
pinkish-orange tinge or brownish to almost
salmon-pink tinge. No soluble pigment.
Nutrient agar: Growth colorless. Aerial
mycelium thin, white. No soluble pigment.
207
Starch agar: Growth same as on synthetic
agar. Aerial mycelium white or white with
light brownish-salmon-pink tinge. Hydrol-
ysis.
Gelatin: Growth yellowish. Aerial myce-
hum in form of white patches. No soluble
pigment. Medium liquefaction.
Potato plug: Growth cream-colored, wrin-
kled. No aerial mycelium. No soluble pig-
ment.
Milk: Growth yellowish, surface
Aerial mycelium white, scant. No soluble
pigment. Coagulation and peptonization.
Blood agar: Growth brownish, wrinkled.
No aerial mycelium. Hemolysis none or
weak.
Antagonistic properties: Produces caryo-
mycin, an antitumor substance.
ring.
78. Streptomyces filipinensis Ammann et
al., 1955 (Ammann, A., Gottheb, D., Brock,
T. D., Carter, H. E., and Whitfield, G. B.
Phytopathology 45: 559-563, 1955).
Morphology: Sporophores form
that vary from open to tightly closed. Spores
spirals
round to oval.
Sucrose nitrate agar: Growth excellent,
hight Aerial mycelium
white, turning gray. Soluble pigment slightly
yellow. cottony,
yellow. Colorless drops of exudate on myce-
lium.
Starch-nitrate agar: Growth excellent.
Aerial mycelium velvety, white, turning
gray. No soluble pigment. Hydrolysis weak.
Glycerol-asparagine agar: Growth excel-
lent. Aerial mycelium white, turning gray.
Soluble pigment slightly yellow.
Nutrient agar: Growth very poor, light
buff. No aerial mycelium. Soluble pigment
brown.
Gelatin: Slow but definite liquefaction,
stratiform type. Soluble pigment brown.
Potato: Growth good. No aerial mycelium.
Soluble pigment purple to black.
Nitrate reduction: Little, if any.
Production of H.S: Positive.
Carbon utilization: Utilizes xylose, arabi-
208
nose, fructose, galactose, sucrose, maltose,
lactose, raffinose, mnulin, mannitol, imositol,
sodium acetate, sodium citrate, sodium
succinate, dextrose, mannose, starch, dex-
trin, and glycerol. Does not utilize rhamnose,
sorbitol, dulcitol, salicin, phenol, m-cresol,
sodium formate, sodium oxalate, sodium
tartrate, or sodium salicylate.
Antagonistic properties: Produces anti-
fungal agent filipin, of the polyene type.
Habitat: Philippine soil.
Type culture: IMRU 3781.
79. Streptomyces fimbriatus (Millard and
Burr, 1926) Waksman and Henrici, 1948
(Millard, W. A. and Burr, 8. Ann. Appl.
Biol. 13: 580, 1926).
Morphology: Sporophores form spirals
with three or more turns. Spores cylindrical
to oval, 1.2 to 0.9 by 0.9 uz.
Sucrose nitrate agar: Growth gray. Aerial
mycelium abundant, white to gray. Soluble
pigment cream-colored.
Glucose-asparagine agar: Growth
good. Aerial mycelium white to mouse-gray.
Nutrient potato agar: Colonies gray to
blackish, flat, raised in center. Aerial myce-
lium a few specks of white. Soluble pigment
very
golden brown.
Potato: Growth mouse-gray. Aerial myce-
lium on dried portions of growth secant, white
to mouse-gray. Pigment around growth
black.
Gelatin: Growth good. Aerial mycelium
white. Liquefaction slow. Soluble pigment
reddish.
Milk: Growth good. No coagulation and
no hydrolysis.
Starch: Positive hydrolysis.
Tyrosinase reaction: Strongly positive.
Nitrate reduction: Positive.
Habitat: Common scab of potatoes.
80. Streptomyces fimicarius (Duché, 1934)
Waksman and Henrici, 1948 (Duché, J.
actinomyces du groupe albus. P. Lechevaler,
Paris, 1934).
Les
THE ACTINOMYCETES, Vol. II
Morphology: Sporophores long, tuft-
forming; no spirals. Spores cylindrical (Hoff-
mann, 1958).
Sucrose nitrate agar: Growth at first color-
less, later yellowish to red-brown; reverse
orange-colored. Aerial mycelium light gray
with yellowish tone. Soluble pigment faint
yellowish.
Glucose-asparagine agar: Growth cream-
colored with whitish aerial mycelium; re-
verse cream-colored to slight ocher.
Nutrient agar: Growth limited, cream-
colored with white aerial mycelium; reverse
yellowish.
Potato: Growth cream-colored to yellow-
ish to dark brown. Aerial mycelium gray.
Soluble pigment reddish-brown.
Gelatin: Punctiform colonies with whitish
aerial mycelium. Soluble pigment reddish.
Liquefaction medium. Melanin-negative.
Milk: Growth colorless, becoming covered
with whitish aerial mycelium. Slow pep-
tonization. Pigment rose, changing to brown-
ish-red.
Starch: Hydrolyzed.
Coagulated serum: Growth cream-colored.
Aerial mycelium whitish. Liquefaction rapid.
Cellulose: No growth.
Tyrosine medium: Growth white, with
yellowish reverse. Soluble pigment yellowish.
Production of HoS: Negative.
Antagonistic properties: Positive.
Krassilnikov (1949)
this organism as a variety of A. chromogenes.
81. Streptomyces flaveolus | (Waksman)
Waksman and Henrici, 1948 (Waksman,
S. A. No. 168. Soil Sci. 8: 134, 1919).
Morphology: Sporophores monopodially
branched. Short, closed and open spirals
temarks: considers
produced on all media. Spores oval to
elliptical, 0.8 by 1.2 uw, covered with long,
fine hair.
Sucrose nitrate agar: Growth light sulfur-
cadmium-yellow. Aerial
to ash-gray. Soluble pig-
vellow, turning
mycelium white
ment yellow.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Malate-glycerol agar: Growth colorless to
cream-colored. Aerial mycelium mouse-gray.
Glucose-asparagine agar: Aerial mycelium
pale gray. Soluble pigment yellowish-green.
Nutrient agar: Growth colorless, glisten-
ing, wrinkled. Aerial mycelium white. Solu-
ble pigment absent or yellow.
Potato: Growth abundant,
cream-colored to yellow. Aerial mycelium
white to pinkish. Soluble pigment absent or
wrinkled,
faint brown.
Gelatin:
spreading. Aerial mycelium white. Liquefac-
tion rapid. Soluble pigment vellowish-brown,
Growth abundant, yellowish,
not melanoid.
Milk: Ring sulfur-yellow. Rapid coagula-
tion and strong peptonization.
Starch media: Growth colorless. Aerial my-
celium light gray. Hydrolysis.
Cellulose: Growth scant.
Invertase: Negative.
Nitrate reduction: Positive.
Production of HoS: Negative.
Antagonistic properties: Some — strains
produce actinomycin.
Habitat: Soil.
Remarks: Several varieties of this organ-
ism have been described. It is sufficient to
mention a culture described by Krassilnikov
as A. rectus, which appears to be a variety
of S. flaveolus. WrassilInikov also believed
that A. krainskia Duché belongs to this
group.
Type culture: IMRU 3319.
82. Streptomyces flavochromogenes (Krain-
sky, 1914) Waksman Henrici, 1948
(Krainsky, A. Centr. Bakteriol. Parasitenk.
Abt. II, 41: 685, 1914).
Morphology: Spores oval, 1.7 wu.
and
Glucose-asparagine agar: Growth yellow.
Aerial mycelium gray. Soluble pigment
brown.
Calcium malate agar: Growth yellow.
Aerial mycelium produced late, white to
oray.
Nutrient agar: Aerial mycelium formed
209
late, at first white, later gray. Soluble pig-
ment brown.
Starch agar: Growth yellow. Aerial myce-
hum white. Weakly diastatic.
Potato: Growth yellow. Aerial mycelium
white. Soluble pigment black.
Gelatin: Colonies yellowish. Slight lique-
faction. Soluble pigment brown.
Cellulose: Growth slow.
Nitrate reduction: Strong.
Tyrosinase: Positive.
Temperature: Optimum
Habitat: Garden soil.
temarks: Krainsky considered this cul-
Bonu:
ture as identical to A. chromogenes Gas-
perini.
Type culture: IMRU 3671.
85. Streptomyces — flavogriseus — (Duché,
1954) Waksman (Duché, J. Les actinomyces
du groupe albus. P. Lechevalier, Paris, 1934).
Morphology: Sporophores long, straight,
with a few curling tips. Spores spherical.
Sucrose nitrate agar: Growth limited,
yellowish, reverse turning black. Aerial
mycehum thin, gray to mouse-gray.
Nutrient agar: thin, cream-
colored. Aerial mycelium thin, white. No
Growth
soluble pigment. Melanin-negative.
Glucose-peptone agar: Growth yellow;
reverse tending to turn dark. Aerial myce-
hum abundant, drab. No
soluble pigment.
Starch agar: Growth very limited, similar
mouse-gray to
to that on sucrose nitrate agar. Hydrolysis.
Potato: Growth abundant, lichenoid.
Aerial mycelium abundant, mouse-gray to
drab with white edge. No soluble pigment.
Gelatin: Growth flocculent, through me-
dium. Liquefaction slow. No soluble pig-
ment.
Milk: Cream-colored ring. No aerial myce-
hum. Peptonization very rapid.
femarks: According to Ettlinger et al.
(1958) the S.
this organism belongs to
fradiae group.
Type culture: IMRU 3322.
210
84. Streptomyces flavoreticult Funaki et al.,
1958 (Funaki, M., Tsuchiya, F., Maeda,
K., and Kamiya, T. J. Antibiotics (Japan)
LLA: 138-142, 1958).
Morphology: Aerial mycelium is long with
many short branches; numerous — small
verticils are produced depending on the
nature of the medium, especially on starch-
ammonium agar.
Sucrose nitrate agar: Growth colorless to
pale yellow. Aerial mycelium white, cottony.
Soluble pigment faint yellow.
Glucose-asparagine agar: Growth pale
yellow to yellowish-brown. Aerial mycelium
yellowish-white to olive-gray. Soluble pig-
ment pale yellow.
Calcium malate agar: Growth pale yellow.
Aerial mycelium pale yellow. No soluble
pigment.
Nutrient agar: Growth yellow to brown.
Aerial mycelium pale yellow to yellowish-
gray. Soluble pigment brown.
Potato: Growth yellow, folded. Aerial
mycelium yellowish white to olive-gray or
olive-yellow. Soluble pigment dark brown.
Milk: Colorless pellicle. Aerial mycelium
white. Soluble pigment brown. Milk coagu-
lated, then peptonized.
Gelatin: Growth yellowish-brown. Aerial
mycelium white to light gray. Soluble pig-
ment brown. Gelatin liquefied.
Starch: Not hydrolyzed.
Antagonistic properties: Produces anti-
biotic virocidin, which possesses antiviral and
antibacterial properties.
Xemarks: Similar to S. reticula and to S.
flavus, differing from the first by the forma-
tion of yellow growth and yellow soluble
pigment and from the second by the forma-
tion of verticils.
85. Streptomyces flavovirens (Waksman,
1919) Waksman and Henrici, 1948 (Waks-
man, 8. A. Soil Sci. 8: 117, 1919).
Morphology: Sporophores coarse, straight,
large
and short, relatively unbranched;
masses of minute tufts; open spirals may be
THE ACTINOMYCETES, Vol. II
produced in certain substrates. Spores
spherical, oval to rod-shaped, 0.75 to 1.0 by
IO Mon 125: pa:
Sucrose nitrate agar: Growth yellowish
with greenish tinge. Aerial mycelium gray.
Soluble pigment greenish-yellow.
Glucose-asparagine agar: Growth — re-
stricted, developing only to a very small
extent into the medium, yellow, turning
black. Soluble pigment golden yellow to
greenish-yellow.
Nutrient agar: Growth yellowish; reverse
dark in center with yellowish zone and
outer white zone.
Potato: Growth sulfur-yellow, wrinkled.
Gelatin: Surface pellicle yellowish-green.
Good liquefaction. Melanin-negative.
Milk: Cream-colored to brownish ring;
coagulation and peptonization.
Starch agar: Growth greenish-yellow,
spreading, developing deep into the medium.
Good hydrolysis.
Invertase: Negative.
Nitrate reduction: Limited.
Production cf HS: Negative.
Cellulose: No growth.
Antagonistic properties: Produces actino-
mycin.
Habitat: Soil.
temarks: Certain forms belonging to this
species, such as A. griseostramineus and A.
olivaceoviridis, have been described by Gause
et al. (1957). Ettlinger et al. (1958) considers
this species as belonging to the S. fradiae
group. Hirsch (1960) considers this organism
as an oligonitrophilic form.
Type culture: IMRU 3320.
86. Streptomyces flavus (Xrainsky, 1914)
Waksman and Henrici, 1948 (kXrainsky, A.
Centr. Bakteriol. Parasitenk. Abt. II, 41:
685, 1914; Waksman, 8S. A. and Curtis,
R. E. Soil Sei. 1: 99, 1916; 8: 71, 1919).
Not A. flavus WKrainsky emend. Krassil-
nikov (1941).
Morphology: Sporophores are long, usu-
== eee
DESCRIPTION OF SPECIES OF STREPTOMYCES
ally no spirals; some open spirals may be
produced. Spores oval, 1.2 yu.
Sucrose nitrate agar: Growth yellow or
sulfur-yellow. Aerial mycelium straw-yellow.
Glucose-asparagine agar: Growth sulfur-
yellow, center shading to brown. Aerial
mycelium white to gray.
Nutrient agar: Growth gray, spreading,
folded. Aerial mycelium white, appears late.
Starch agar: Growth cream-colored with
pink tinge. Hydrolysis marked.
Potato: Growth yellow. Aerial mycelium
gray. Melanin-negative.
Gelatin: Growth in form of small, yellow-
ish masses on surface. Rapid liquefaction.
Melanin-negative.
Milk: Rapid coagulation and peptoniza-
tion.
Sucrose inversion: Negative.
Nitrate: No reduction.
Cellulose: Growth poor.
Temperature: Optimum 25°C.
Antagonistic properties: Some strains
produce actinomycin and certain other
antibiotics.
Habitat: Soil.
Remarks: Represents a large group of
species, as shown previously (Chapter 3).
Above description is based largely upon that
given by Krainsky. According to Ettlinger
et al. (1958), this organism does not form
any spirals (as found also by Waksman and
Curtis) and is related to S. olzvaceus.
Type culture: IMRU 3321.
87. Streptomyces flocculus (Duché, 1934)
Waksman and Henrici (Duché, J.
actinomyces du groupe albus. P. Lechevalier,
Paris, 1934).
nitrate
Les
Sucrose agar: Growth cream-
later covered with white aerial
mycelium. No soluble pigment.
colored,
Glucose-asparagine agar: Growth limited,
cream-colored, only slightly raised above the
surface of the medium; occasionally abun-
dant growth produced with white aerial
mycelium, colorless on reverse side.
PA
Nutrient agar: Growth cream-colored,
later covered with white aerial mycelium.
No soluble pigment.
Potato: Growth punctiform. Aerial myce-
lum white. Soluble pigment faint, yellowish.
Gelatin: Growth limited.
slow. Melanin-negative.
Liquefaction
Milk: Growth rose-colored. Peptonization
slow.
Coagulated serum: Growth cream-colored.
Aerial mycelium fine, white. Liquefaction
slow.
Tyrosine medium: Growth whitish. No
soluble pigment.
Production of HS: Negative.
Remarks: Belongs to the S. albus series.
88. Streptomyces fradiae (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, 8. A. and Curtis, R. E. Soil Sei.
1: 99-134, 1916; 8: 90, 1919).
Morphology: | Sporophores — branched
monopodially, straight or flexible, but no
true spirals. On certain media, such as
glycerol agar, spirals are formed. Ettlinger
et al. (1958) found open spirals. Spores oval
to rod-shaped, 0.5 by 0.7 to 1.25 uw, smooth
(Fig. 39).
Sucrose nitrate agar: Growth smooth,
spreading, colorless, or pale yellow-orange.
Aerial mycelium thick, cottony, seashell-
pink. No soluble pigment.
Malate-glycerol
Aerial mycelium seashell-pink.
agar: Growth orange.
Glucose-asparagine Growth re-
stricted, glossy, buff-colored, lichenoid mar-
agar:
gin. Aerial mycelium appears late, seashell-
pink.
Nutrient agar: Growth restricted, yellow-
ish, becoming orange-yellow to buff. No
aerial mycelium. No soluble pigment.
Potato: Growth restricted, orange-colored.
Aerial mycelium white to rose or pink. Solu-
ble pigment absent or faint brown.
Gelatin: Growth dense, cream-colored to
brownish. Aerial mycelium white. Gelatin
liquefied. No soluble pigment.
FiGuRE 39. Sporophores of S. fradiae (Prepared
by H. Lechevalier of the Institute of Microbiol-
ogy).
Starch media: Growth spreading, colorless.
Aerial mycelium seashell-pink. Good dia-
static action.
Milk: Cream-colored ring; coagulation
and rapid peptonization.
Nitrate: Varied reduction.
Cellulose: No growth in solution, fair
growth on plates.
Production of H.S: Negative.
Invertase: None.
Antagonistic properties: Highly antag-
antibacterial agent,
neomycin, and an antifungal agent, fradicin.
Habitat: Soil.
temarks: A
onistic. Produces an
this
organism have been isolated from various
number of strains. of
soils (see, for example, S. decaris described as
No. 3719, in Waksman et al., 1958). Some
vary in their pigmentation, rate of gelatin
liquefaction, and antibiotic production; A.
longissimus IKrassilnikov is one such typical
strain. Some strains are able to produce
antiviral substances, as In the case of luridin,
produced by a strain of S. fradiae designated
as A. luridus by Wrassilnikov et al. Gause
et al. (1957) described a_ spiral-producing
variety of S. fradiae under the name spiralis.
Several other such strains have been isolated
by Waksman and Lechevalier, Umezawa,
and many others. Ettlinger et al. (1958)
claimed that S.
rochei, S. filipinensis, S.
2 THE ACTINOMYCETES, Vol. II
coelicolor, S. flavogriseus, S. tyrosinaticus, S.
violaceus, and S. violaceoruber belong to this
group; this claim cannot be accepted on the
basis of evidence submitted in the descrip-
tions of these organisms. The characteristics
of the species are that it is nonchromogenic,
strongly proteolytic, and produces the
characteristic seashell-pink aerial mycelium
on various synthetic media; on organic
media, orange-colored growth is produced
without any aerial mycelium.
Kor
Type culture: IMRU 3535.
89. Streptomyces fragilis Anderson et al.,
1956 (Anderson, L. E., Ehrlich, J., Sun,
S. H., and Burkholder, P. R. Antibiotics &
Chemotherapy 6: 100, 1956).
Morphology: Aerial hyphae simple or
branched, usually in small clusters; short,
straight, or slightly curved, with bent or
curved tips and occasional short spirals.
Spores spherical to ovoid, 0.8 to 1.5 by 1.0
to 2.0 p.
Calcium malate agar:
Aerial mycelium light brown.
Growth sparse.
Glycerol-asparagine agar: Growth sparse,
colorless to light yellow. Aerial mycelium
hight yvellow-pink.
Starch-ammonium sulfate agar: Growth
vellow to yellow-orange to orange-brown.
Aerial mycelium light vellow-pink to lght
brown.
Nutrient agar: Growth yellow to yellow-
orange to brown. Aerial mycelium white to
light yellow-pink. Melanin-negative.
Glucose-tryptone agar: Growth yellow to
yellow-orange to brown. Aerial mycelium
light vellow-pink, occasionally pink to gray-
pink.
Gelatin: Liquefaction slow. No soluble
pigment.
Litmus milk: Slow peptonization.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Carbon utilization: Utilizes L-arabinose,
p-cellobiose, dextrin, b-galactose, glucose,
p-maltose, starch, trehalose, and p-xylose.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Does not utilize aesculin, adonitol, cellulose,
citrate, dulcitol, glycerol, 7-inositol, inulin,
p-levulose,
nose, melezitose, melibiose, pb-raffinose, L-
p-lactose, D-mannitol, b-man-
rhamnose, salicin, p-sorbitol, succinate, or
sucrose.
Antagonistic properties: Produces a sub-
that
anticancer properties.
Source: Argentine soil.
femarks: Closely resembles S. fradiae.
stance, azaserine, possesses certain
A detailed comparison between this and
closely related organisms has been made by
Anderson ef al. (1956).
Type culture: IMRU 3732, NRRL 2424.
90. Streptomyces fulvissimus (Jensen,
1930) Waksman and Henrici, 1948 (Jensen,
H. L. Soil Sei. 30: 66, 1930).
Morphology: Sporophores short, straight,
often trifurcated. Slightly wavy, but no true
spirals. Spores oblong, smooth, 1.0 to 1.2
Dyaleatonh osu. blind).
Sucrose nitrate agar: Growth light golden,
later deep orange to red-brown. Aerial myce-
lium scant, white, later grayish-brown.
Soluble pigment bright golden to orange.
Glycerol-asparagine agar: Growth golden
to dark brown. Aerial mycelium white to
light Soluble pigment
golden to orange.
Nutrient agar: Growth wrinkled, deep
cinnamon-brown.
red-brown. No aerial mycelium. Soluble pig-
ment brownish-yellow. Melanin-negative.
Potato: Growth wrinkled,
Aerial mycelium absent or white. Soluble
rust-brown.
pigment gray to faint lemon-yellow.
Gelatin: Growth yellowish-brown to red-
brown. No aerial mycelium. No soluble pig-
ment. Rapid liquefaction.
agar: Growth
brown. Aerial mycelium hydrolyzed, smooth,
lead-gray. Soluble pigment dull yellow to
Starch-caseim vellowish-
orange starch.
Production of HS: Positive.
Antagonistic properties: Produces valino-
mycin.
Habitat: Very common in soil.
Remarks: The characteristic golden pig-
ment is formed in nearly all media, but
becomes most typical and attains its greatest
brightness in synthetic agar media. It has
indicator properties, turning red in strongly
acid solutions. The species is easily recog-
nized on agar plates by its bronze-colored
colonies, surrounded by halos of bright yel-
low pigment.
This species was believed to be identical
with the culture described by Millard and
Burr (1926) as A. flavus. The last name is
invalid, however, since the culture could be
readily distinguished from the S. flavus of
Krainsky (1914, emend. Waksman
Curtis, 1916) Waksman and Henrici.
Type culture: IMRU 3665.
and
91. Streptomyces fumosus (Krassilnikoyv,
mycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 58, 1941).
Morphology: Sporophores straight. Spores
cylindrical, later round, 1.5 to 2.0 by 0.7 u.
Sucrose nitrate agar: Growth dark brown,
pigment insoluble. Aerial mycelium well
developed, cottony, dust-colored, occa-
sionally gray-white.
Nutrient agar: Growth dark brown. Aerial
mycelium white. Soluble pigment brown.
Potato: Aerial mycelium absent or only
faint, dark gray. Melanin-negative.
Gelatin: Liquefaction medium.
Milk: No coagulation; slow liquefaction.
Soluble pigment dark brown to almost black.
Starch: Good hydrolysis.
Cellulose: No growth.
Sucrose: Inversion weak.
Antagonistic properties: None
Habitat: Soil.
92. Streptomyces fungicidicus Okami et al.,
1954 (Okami, Y., Utahara, R., Nakamura,
S., and Umezawa, H. J. Antibiotics (Japan)
7A: 100-101, 1954).
Morphology: Aerial mycelium produces
214
numerous spirals on most synthetic media.
Spores spherical to oval.
Glycerol nitrate agar: Growth colorless.
Aerial mycelium white to grayish.
Glucose-asparagine agar: Growth color-
less. Aerial mycelium white to grayish. Solu-
ble pigment sometimes yellowish.
Calcium malate agar: Growth colorless to
yellowish. Aerial mycelium white to grayish.
Soluble pigment of some strains pink; later
disappears.
Nutrient agar: Soluble pigment absent or
slightly yellowish-brown. Melanin-negative.
Starch agar: Growth colorless to yellowish.
Aerial mycelium white to grayish. Strong
hydrolysis.
Potato: Growth erayish.
Aerial mycelium absent or white to grayish.
A deep brown soluble pigment around the
growth may be produced.
Milk: Growth colorless to cream-colored.
Soluble pigment absent or shghtly brown.
Coagulation and peptonization weak.
Gelatin: Growth colorless — to
colored. Soluble pigment absent or faint
yellowish to
crealm-
brown. Positive liquefaction.
Nitrate: No, or doubtful reduction.
Tyrosinase: Some strains positive.
Antagonistic properties: Produces a poly-
ene-type antifungal substance, fungicidin.
femarks: Two groups of this species were
recognized; Group A produces nonspiral-
forming sporophores; aerial mycelium white;
violet pigment on potato. It is thus differ-
entiated from Group G, described above.
93. Streptomyces fuscus (Sdhngen and Fol,
1914) Waksman (Séhngen, N. L. and Fol,
J. G. Centr. Bakteriol. Parasitenk. Abt. IT,
40: 89-98, 1914).
Morphology: According to Irassilnikovy,
the organism forms short straight sporo-
phores arranged in fascicles or clusters in the
form of brushes.
Agar media: Growth irregular, dry, color-
less to stone-red. Aerial mycelium initially
white, later becoming dark brown. Spores
THE ACTINOMYCETES, Vol. II
rose-colored. Some strains excrete a brown
substance in protein media.
Carbohydrates: Shght decomposition;
even glucose is assimilated with difficulty.
Carbon utilization: Best are
calcium salts of various organic acids, rang-
ing from malic and citric to stearate and
palmitate. Formate not utilized.
Nitrogen utilization: Ammonium chloride
and asparagine, nitrate, and peptone assimi-
lated with difficulty.
Paraffins: Assimilated.
Rubber: Brown-red growth. Rubber de-
composed.
Temperature: Optimum 33°C; maximum
°C. Destroyed in 5 minutes at 65°C.
Habitat: Soil.
sources
lod
or
94. Streptomyces galbus Frommer, 1959
(Frommer, W. Arch. Mikrobiol. 32: 195,
1959).
Morphology: Sporophores monopodially
branched, ending in spirals with 3 to 8 turns.
On some media, certain strains produce long,
straight, slightly branched aerial hyphae,
with short side branches.
Glycerol nitrate agar: Growth abundant;
Aerial
mouse-gray, or
reverse yellow to yellow-green.
mycelium
green-gray. Soluble pigment golden yellow,
cream-colored,
later turning green-yellow.
Glucose-asparagine agar: Growth weak,
crusty, reverse light yellow or green-vellow,
later turning brown. Aerial mycelium thin,
powdery, with white spots.
Soluble pigment yellowish to yellow-green.
Calcium malate agar: Growth yellow to
greenish-yellow. Aerial mycelium white to
brownish-gray. Soluble pigment yellow to
mouse-gray
vellow-green.
Nutrient agar: Growth thin, brown. No
aerial mycelium. Soluble pigment brown.
Starch-KNO; agar: Growth yellow. Aerial
mycelium white to white-gray. Soluble pig-
ment yellow. Slow hydrolysis of starch.
Potato: Growth heavy, yellow to reddish-
brown. Aerial mycelium powdery, white,
DESCRIPTION OF SPECIES OF STREPTOMYCES
mouse-gray to green-gray. Soluble pigment
black.
Gelatin: Growth abundant. Aerial myce-
lum yellow. Soluble pigment dark brown.
Slow liquefaction.
Milk: No coagulation; slow peptonization.
Cellulose: Growth good.
Antagonistic properties: Produces actino-
mycin.
Remarks: Related to S. vzridochromogenes,
S. flavochromogenes, and S. viridoflavus. It
was also said to be related to S. parvullus. A
variety of this species designated as achro-
mogenes, not producing any melanin pig-
ment, was also described.
95. Streptomyces galilaeus Ettlinger et al.,
1958 (Ettlnger, L., Corbaz, R., and Hiitter,
R. Arch. Mikrobiol. 31: 356, 1958).
Morphology: Sporophores monopodially
branched, with open, regular spirals. Spores
smooth (Pl. II k).
Glycerol nitrate agar: Substrate growth
at first light carmine, later carmine-red.
Aerial mycelium white-gray. No soluble pig-
ment.
Glucose-asparagine agar: Growth thin,
white-vellow, later red. Aerial mycelium
ash-gray. No soluble pigment.
Calcium malate agar: Growth thin, white-
yellow, later red. Aerial mycelium ash-gray.
No soluble pigment.
Starch agar: Growth carmine-red. Aerial
mycelium white-gray. Limited hydrolysis.
Potato: Growth brownish-yellow. Aerial
mycelium ash-gray. Soluble pigment limited,
chestnut-brown. Melanin-positive.
Gelatin: Surface growth light red to light
brown. Aerial mycelium sparse, grayish-
white. Soluble pigment reddish-brown to
dark brown. Liquefaction trace. Melanin-
positive.
Milk: Pellicle thick, light brown. Aerial
mycelium ash-gray. Limited coagulation, no
peptonization.
Antagonistic properties: Positive.
Habitat: Soil.
215
96. Streptomyces galtiert (Goret and Jou-
bert, 1951) Waksman (Goret, P. and Jou-
bert, L. Ann. parasitol. humaine et comparée
26: 118-127, 1951).
Morphology: Two types of colonies are
produced on agar: one small, flat, regular,
white; the other thick, irregular,
yellowish. Sporophores form spirals. Spores
oval, 0.8 to 1.5 by 0.8 up.
large,
Sucrose nitrate agar: Growth limited.
Aerial mycelium powdery, white. No soluble
pigment.
Nutrient agar: Growth poor, thin, yellow-
ish. Aerial mycelium powdery, white. Solu-
ble pigment brown.
Peptone agar: Growth limited, cream-
colored. Aerial mycelium powdery, white.
Soluble pigment very slight, brown-reddish.
Starch agar: Growth thin. Aerial myce-
lum powdery, white. No soluble pigment.
Potato: Punctiform colonies growing to-
gether as thick crust, orange-reddish in
color. Aerial mycelium limited, white, ap-
pearing very slowly. No soluble pigment.
Gelatin: Growth poor, flaky, white.
Liquefaction limited.
Milk: Growth Aerial
At 25°C no coagulation; at
mycelium
Bye ©.
coagulation after 20 days; no peptonization.
slow.
white.
Nitrate reduction: Positive.
Production of H.S: Negative.
Source: Dog septicemia (thoracic, ab-
dominal, and brain lesions).
femarks: Said to be pathogenic for
guinea pig and rabbit. Culture grown in the
laboratory not pathogenic for dogs.
97. Streptomyces gardnert (Waksman and
Henrici) nov. comb. (Gardner, A. D. and
Cham: i. Brit. J.. Exptl. Pathol: 232: 123.
1942; Waksman, S. A., Horning, E. S.
Welsch, M., and Woodruff, H. B. Soil Sci.
04: 289, 1942).
Morphology: When grown on oatmeal
agar, aerial mycelium thin, largely at edge
of growth, consisting of short, straight to
216
wavy sporophores, often produced in clus-
ters; no spirals.
Glucose-asparagine agar: Growth brown-
ish, lichenoid, with wide cream-colored
edge; reverse yellowish. Aerial mycelium
white to grayish, gradually covering surface.
No soluble pigment.
Nutrient agar: Growth
elevated, lichenoid, doughy consistency. No
Soluble pigment faint
cream-colored,
aerial mycelium.
brownish.
Potato: Growth reddish-
brown. No aerial mycelium. Pigment around
barnacle-like,
erowth grayish-brown.
Gelatin: Surface ring cream-colored. Liq-
uefaction medium. Soluble pigment deep
brown, gradually diffusing through liquefied
portion.
Tryptone broth: Growth occurs in form
of small pellets at the base of the flask; later,
a thin surface pellicle is produced. Soluble
pigment produced slowly, black.
Temperature: Good growth at 25°C; slow
growth at 37°C.
Antagonistic properties:
synthetic and organic media
Produces on
an antibiotic,
proactinomyein, active against bacteria.
Source: Isolated as an air contaminant.
Remarks: This species was first classified
as a Nocardia (Bergey’s Manual, 7th ed.).
Recent evidence, comprising both cultural
(Cummins and
and chemical
Harris, 1958), suggests its transfer to the
properties
genus Streptomyces. It is closely related to
S. aureofaciens.
Type culture: IMRU 3834.
98. Streptomyces garyphalus Harris et al.,
1955 (Harris, D. A., Ruger, M., Reagan,
M. A., Wolf, F. J., Peck, R. L., Wallick, H.,
and Woodruff, H. B. Antibiotics & Chemo-
therapy 5: 183-190, 1955).
Morphology: Sporophores straight, with-
out spirals. Spores rod-shaped, 0.8 to 1.1 by
Lat to: 129 iq:
Growth colorless.
Sucrose nitrate agar:
THE ACTINOMYCETES, Vol. II
Aerial mycelium grayish-white. No soluble
pigment.
Glucose-asparagine agar: Growth color-
less. Aerial mycelium white. No soluble pig-
ment.
Modified glucose-asparagine agar: Growth
powdery, pinkish-white, reverse buff. Aerial
mycelium seashell-pink. No soluble pigment.
Calcium malate agar: Growth colorless
No aerial mycelium.
Nutrient agar: Growth colorless. Aerial
mycelhum grayish-white. Soluble pigment
faint brown.
Yeast extract-glucose agar: Growth excel-
lent. Aerial myceluum = grayish-white, be-
coming pinkish-gray and _ finally seashell-
pink. Soluble pigment faint brown.
Starch-tryptone agar: Growth good. Aerial
mycelium gray. Soluble pigment dark brown.
Peptone-glucose agar: Growth cream-
colored. Aerial mycelium grayish-white, be-
coming pink. Soluble pigment faint brown.
Starch agar: Growth Aerial
mycelium white to gray. Hydrolysis. Soluble
pigment faint brown.
excellent.
Gelatin: Grayish-white ring and sub-
merged pellicle. Pigmented layer dark
brown; becomes greenish-brown when
shaken. Medium liquefaction.
Milk: Faint grayish-white tinge. Slow
peptonization turning dark purple at first
and later brownish-purple. Reaction acid,
pH 6.4.
Nitrate reduction: Strong.
Potato: Growth heavy, wrinkled. Aerial
mycelium grayish-black. Potato darkened.
Cellulose: No decomposition.
Antagonistic properties: Produces an anti-
biotic, pb-4+-amino-3-isoxazolidone (novobio-
cin).
Habitat: Soil.
99. Streptomyces gedanensis — (Lohlein,
1909) Miller, 1950 (Lohlein, M. Z. Hyg.
Infektionskrankh. 63: 1-16, 1909; Miller, R.
Medizinsche Mikrobiologie, 4th ed. 1950,
Urban & Schwarzenberg, Miinich, p. 294).
DESCRIPTION OF SPECIES OF STREPTOMYCES
Morphology: Aerial mycelium consists of
short, gnarled hyphae. Spores short, oval to
spherical.
Synthetic agar: Growth dark to almost
black, with dark reverse. Aerial mycelium
abundant, mouse-gray. No soluble pigment.
Nutrient agar: Growth thin, colorless. No
aerial mycelium. No soluble pigment.
Glucose agar: Growth cream-colored, be-
coming black with light margin. Aerial myce-
lum abundant, mouse-gray.
Potato: Growth lichenoid, cream-colored
to brownish. No aerial mycelium. No soluble
pigment.
Gelatin: Growth thin, flaky. No soluble
pigment. Rapid liquefaction.
Milk: Surface
peptonization.
Starch media: Growth yellowish to cream-
colored. Aerial mycelium light gray. Hy-
ring cream-colored. No
drolysis strong.
Nitrate reduction: Negative.
Production of HS: Negative.
Source: Sputum of patient with chronic
lung disease.
Type culture: IMRU 3417.
100. Streptomyces gelaticus (Waksman,
1919) Waksman and Henrici, 1948 (Waks-
man, 8S. A. Soil Sci. 8: 165, 1919).
Synonym: Streptomyces hepaticus.
Morphology: Sporophores produce open
spirals. According to Anderson ef al. (1956),
the organism does not produce spirals.
Sucrose nitrate agar: Growth colorless,
spreading, chiefly deep into the medium.
Aerial mycelium thin, white, turning grayish.
Nutrient agar: Growth only on surface,
wrinkled, cream-colored.
Glucose agar: Growth abundant, spread-
ing, white.
Potato: Growth abundant, much wrin-
kled, greenish, becoming black with yellow-
ish margin.
Gelatin: Produces flaky, cream-colored
sediment. Good liquefaction.
Pa ef
Milk:
peptonization.
Starch: Growth thin, spreading, cream-
colored. Hydrolysis.
Nitrate reduction: Positive.
Production of H.S: Negative.
Temperature: Optimum 25°C.
Pinkish ring. Coagulation and
Antagonistic properties: Produces elaio-
mycin.
Habitat: Soil.
temarks: Various. related
been described by Gause ef al. (1957); these
include A.
forms have
griseorubens, A. rubiginosus, and
KrassilInikov (1959)
siders this organism as belonging to the S.
A. atroolivaceus. con-
albus group.
Type culture: IMRU 3323.
101. Streptomyces glaucus (Lehmann and
Schiitze emend. Krassilnikov, 1941) Waks-
man (KrassiInikov, N. A. Actinomycetales.
Izvest. Akad. Nauk. SSSR, Moskau, p. 46,
1941).
Morphology: Sporophores form compact
spirals with 3 to 5 turns. Spores oval to
spherical, 1.0 by 0.8 uy.
Sucrose nitrate agar: Growth colorless:
soluble pigment brown. Aerial mycelium at
first white, then becoming bright green.
Nutrient heavy. Aerial
mycelium green.
Potato: Growth heavy. Aerial mycelium
Growth
agar:
velvety, green.
Gelatin: Liquefaction slow. Melanin-nega-
tive.
Milk:
coagulation by some strains.
Starch: Hydrolysis rapid.
Cellulose: Growth good.
Nitrate reduction: Positive.
Peptonization slow, with prior
Sucrose: Poor inversion.
Paraffin: Growth good.
Antagonistic properties: All — strains
strongly antagonistic.
Habitat: Soil.
temarks: Numerous cultures belonging
to this organism or closely related to it have
218
been described under a variety of different
names. It 1s sufficient to mention S. caelestis,
which produces an antibiotic, celesticetin,
described by DeBoer et al. (1954) and A.
glaucescens, together with a variety badius,
described by Gause et al. (1957).
102. Streptomyces globisporus (kXrassilni-
kov, 1941) Waksman (Krassilnikov, N. A.
Actinomycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 48, 1941).
Morphology: Sporophores
wavy, often gathered in clusters or tufts; no
spirals. Spores oval (1.2 to 1.4 by 1.8 to
2.0 uw) or spherical (0.9 to 1.4 yu).
Starch-KNO; agar: Growth abundant,
colorless. Aerial mycelium light yellow to
greenish-yellow with pinkish No
soluble pigment.
Glucose-peptone agar: Growth colorless
or greenish. Aerial mycelium creamy, seldom
greenish-yellow. Soluble pigment absent or
faint yellowish.
Gelatin: Rapid liquefaction. Soluble pig-
ment absent or hight brownish.
Potato: Growth colorless or
Aerial mycelium
brownish or colorless.
Milk: No coagulation; rapid peptoniza-
tion.
Starch: Weak hydrolysis.
Invertase: None.
Nitrate: Reduced to nitrite.
Cellulose: No or poor growth.
Some
straight or
tinge.
brownish.
greenish-yellow. Plug
Antagonistic properties: straims
suppress gram-positive bacteria.
Habitat: Soil.
temarks: Krassilnikov recognized several
substrains of the species on the basis of
milk coagulation, proteolysis, and pigmenta-
tion of aerial mycelium. It is sufficient to
mention A. globisporus vulgaris, A. globis-
porus griseus, A. globisporus lactis, A. globis-
porus diastaticus, A. globisporus flaveolus, A.
globisporus circulatus, A. globisporus scabies,
and A. globisporus albus. This heterogeneous
THE ACTINOMYCRETES, Vol. II
collection is most unfortunate, since these
ce 9)
species”? show differences in color of aerial
mycelium, in formation of soluble pigments,
etc.
Krassilnikov (1949) considered the strep-
tomycin-producing organism as a_ variety
of this species, designating it at first as A.
globisporus streptomycini, and later as A.
streptomycin, A. Krainsky
distinguished from this species on the basis
of the fact that the sporophores of the latter
exhibited spiral formation. This again was
the cause of much confusion in nomenclature
griseus was
of the streptomycin-producing organism in
the literature of the Soviet Union.
Later, Krassilnikov (1958) divided the A.
globisporus group, on the basis of antago-
nistic effects, Into a number of subgroups,
including A. vulgaris, A. toxicus, A. levoris,
A. bacillaris, A. fluorescens, A. raffinosus, A.
longisporus, and A. grisinus.
Gause et al. (1957) described A.
porus in the series “‘helvolus,’’ comprising
globis-
the S. griseus group; they also listed several
additional forms belonging to S. globisporus
under the names A. caucasicus and A.
cyanofuscatus. The above description is
based upon the comparison made by Gause
et al. of six cultures and Krassilnikov’s
authentic strain.
103. Streptomyces globosus (Xrassilnikov,
1941) Waksman (Krassilnikov, N. A. Actino-
mycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 58, 1941).
Morphology: Sporophores straight, short,
slightly branched, wavy. Spores spherical to
oval.
Agar media: Substrate growth brown to
dark brown. Aerial mycelium dark gray,
velvety. Soluble pigment dark brown.
Gelatin: Weak liquefaction. No soluble
pigment. Melanin formation questioned by
Hoffmann (1958).
Potato: Soluble pigment red-brown (Hoff-
mann, 1958).
DESCRIPTION OF
Milk: Questionable
peptonization.
Starch: Hydrolysis.
Cellulose: ¢
No inversion.
: Positive.
coagulation; good
wood growth.
Sucrose:
Production of H.S
Antagonistic properties: No activity.
Habitat: Soil, food products, potatoes.
Type culture: IMRU 3736
104. Streptomyces gougeroti (Duché, 1934)
Waksman and Henrici, 1948 (Duché, J. Les
actinomyces du groupe albus. P. Lechevalier,
Paris, 1934)
Morphology: Aerial hyphae short, gnarled.
Spores oval.
Glucose slow as
punctiform with
smooth edge. No aerial mycelium. No soluble
agar: Growth
cream-colored
nitrate
colonies;
pigment.
Glucose-asparagine agar: Growth colorless
to yellowish. Aerial mycelium thin, white.
No soluble pigment.
Nutrient agar: Growth »am-colored
with brownish reverse. Aerial mycelium
thin, white. Soluble pigment faint yellowish.
Potato: Growth slow, greenish tinged.
Aerial mycelium thin, white. No soluble pig-
ment.
Gelatin: Surface growth heavy, cream-
colored. Aerial mycelium thin, white. Lique-
faction strong. Melanin-negative.
Milk: Growth cream-colored.
celium thin, white. Peptonization rapid.
Coagulated serum: Growth cream-colored.
Aerial mycelium white. Liquefaction rapid.
Starch: Hydrolysis rapid.
Nitrate reduction: Negative.
Production of H.S: Negative.
properties: Active
Aerial my-
Antagonistic against
fungi.
Remarks:
intermediate
abundant aerial mycelium, and S. almquist?,
with its very scant aerial mycelium.
Type culture: IMRU 3590.
This culture is believed to be
between S. albus, with its
SPECIES OF
STREPTOMYCES 219
105. Streptomyces gracilis Eee and
Burr, 1926) Waksman (Millard, W. A. and
Burr, S. Ann. Appl Biol. 13: 580, ae
Morphology:
Spores oval or spherical, 0.8 to 0.9 by 0.8 yp.
fern-like,
gray to
Sporophores form. spirals.
nitrate Growth
pale gray. Aerial mycelium scant,
buff. Soluble pigment cream-colored.
Nutrient potato agar:
buff to dark brown or almost black. Aerial
Sucrose agar:
Growth vinaceous-
mycehum gray. Soluble pigment light golden
brown.
Potato:
gray to buff
Gelatin:
white. Liquefaction
pink to dark golden brown.
Milk: Surface growth good. Aerial myce-
lium white in the form of a ring and specks
Coagulation slow, followed by
Aerial mycelium abundant, olive-
f. Plug pigmented light brown.
Growth gray. Aerial mycelium
rapid. Soluble pigment
on surface.
rapid peptonization.
Starch: Positive hydrolysis.
Nitrate reduction: Positive.
Tyrosinase reaction: Negative.
Temperature: Grows well at 37.5°
Habitat:
106. Streptomyces
Potato scab.
grisenus Waksman
(Reynolds, D. M. and Waksman, S. A.
J. Bacteriol., 55: 739-751, 1948; Okami, Y.
J. Antibiotics (Japan) 3: 95-97, 1950).
Morphology: Straight sporophores pro-
duced clusters or tufts, without spirals.
Spores rod-shaped, 1.0 to 1.8 by 0.8 to 1.0 xu.
nitrate agar: Substrate growth
-am-colored to brownish.
Sucrose
wrinkled, reverse cr
Aerial mycelium white to
with light greenish tinge (lesser tendency to
cream-colored
grass-green coloration, more of a cream-
color). No soluble pigment.
Starch agar: Colorless
Aerial mycelium grayish-olive.
to cream-colored
growth. Hy-
drolysis rapid.
Potato: Growth wrinkled, yellowish-white.
Aerial with olive
tinge.
mycelium grayish-white
220
Gelatin: Growth cream-colored with
brownish tinge. Aerial mycelium absent, or
scant, white. Liquefaction rapid.
Milk: Growth cream-colored. Coagulation
and peptonization.
Tyrosine agar: No pigment produced.
Nitrate reduction: Positive.
Production of HS: Negative.
Carbon utilization: Okami (1950) re-
ported that the grisein-producing organism
GS. griseinus) grows more readily in synthetic
media contaming and
sucrose than the streptomycin-producing S.
griseus. According to Benedict et al. (1955),
the former utilizes xylose, L-arabinose, and
glucose, glycerol,
rhamnose, but S. griseus utilizes only xylose.
Phage sensitivity: Not sensitive to phages
effective against S. griseus strains.
Pigments: No soluble pigments on calcium
malate or succinate media, whereas S. gr7-
seus forms green and yellow pigments on
these media, according to Benedict and Lind-
enfelser (1951).
Antagonistic properties: Produces the an-
tibiotic grisein. Albomycin, produced by A.
subtropicus (Gause, 1955), is an identical or
closely related compound (Waksman, 1957;
Thrum, 1957).
femarks: S. griseus and S. griseinus show
other striking differences. There are some
close resemblances between these and the
viomycin-producing cultures. Thus, on tyro-
sine-starch agar, certain S. griseus strains
form a dark pigment in the agar, whereas S.
griseinus strains resemble the viomycin-pro-
ducing cultures by not forming this pigment.
S. griseinus and the viomycin group grow
well on NaNO;, but S. griseus utilizes this
compound poorly. A. subtropicus, described
by Jkudrina and Kochetkova (1958), is
closely related to, if not identical with SS.
Griseinus.
Type culture: IMRU 3478.
107. Streptomyces griseobrunneus Waks-
man, 1919 (Waksman, 8. A. Soil Sei. 8: 125
127-1919):
THE ACTINOMYCETES, Vol. II
Morphology: Sporophores usually straight
on most media; often a few short, open
spirals are formed; tufts are produced on
certain media. Spores oval-shaped.
Sucrose nitrate agar: Growth cream-col-
ored to yellowish-brown. Aerial mycelium
appears early; powdery, olive-buff to water-
green. No soluble pigment.
malate Growth
cream-colored. Aerial mycelium water-green
in color.
Glycerol-caletum
agar:
Glucose-asparagine agar: Growth yellow-
ish-brown. Aerial mycelium pale olive-buff.
No soluble pigment.
Nutrient agar: Growth cream-colored, be-
coming brown. Aerial mycelium abundant,
white. Soluble pigment brown.
Starch agar: Growth cream-colored to yel-
lowish. Aerial mycelium white. Good hydrol-
ysis.
Ege media: Growth cream-colored with
455
brownish tinge. Aerial mycelium olive-buff.
Soluble pigment purple.
Potato: Growth brownish. Aerial myce-
lium white, turning olive-buff. Soluble pig-
ment faintly brown.
Gelatin: Growth cream-colored, turning
brown. Aerial mycelium white. Soluble pig-
ment deep brown. Medium liquefaction.
Milk: No coagulation; rapid peptoniza-
tion.
Nitrate reduction: Positive.
Sucrose: No inversion.
Cellulose: Good growth.
Habitat: Sewage.
Yemarks: This organism had been de-
scribed by Waksman (1919) as Actinomyces
218, but never named before. It was said to
be closely related to S. griseus, differing from
it by lesser proteolysis and production of a
brown pigment on protein media.
Type culture: IMRU 3068.
108. Streptomyces griseocarneus Benedict
et al., 1951 (Benedict, R. G., Lindenfelser,
L. A., Stodola, F. H., and Traufler, D. H.
J. Bacteriol. 62: 487-497, 1951; see also
DESCRIPTION OF SPECIES OF STREPTOMYCES 221
Grundy, W. E. Antibiotics & Chemother-
apy 1: 309-317, 1951).
Morphology: Sporulation occurs best on a
carbon-free salt agar, to which 0.5 per cent
soluble starch has been added. Sporophores
straight, forming no spirals. Spores coccoid
toroveal lletoslG2by O27 told.L a:
Agar media: Aerial mycelium on some
media powdery, becoming gray, but no spor-
ulation. When sporulation occurs the myce-
lium becomes light pink.
limited,
nitrate Growth
white. Aerial mycelium white, no sporula-
Sucrose agar:
tion.
Glucose-asparagine agar: Growth moder-
ate. Aerial mycelium powdery, white, no
sporulation.
Caletum malate agar: Growth moderate,
white. Aerial mycelium white, no sporula-
tion.
Nutrient agar: Growth moderate, cream-
colored. No aerial mycelium. Soluble pig-
ment light yellow-brown.
Oatmeal agar: Growth luxuriant, brown.
Aerial mycelium abundant, fluffy, white; no
sporulation. No soluble pigment.
Potato: Growth cream-colored. Aerial my-
celium gray. Soluble pigment hght brown,
turning dark brown.
Gelatin: Growth cream-colored to brown.
Rapid liquefaction. Soluble pigment dark
brown.
Milk: Growth dark brown to black. No
coagulation; rapid peptonization. Soluble
pigment brown.
Starch: Hydrolysis.
Carbon — utilization:
starch, glycerol, calcium malate, and sodium
Glucose, dextrin,
succinate rapidly utilized. Mannose, mal-
tose, inositol, and sodium acetate utilized
slowly. Xylose, galactose, sorbose, sucrose,
cellobiose, melibiose, lactose, mannitol, sor-
bitol, sodium citrate, and potassium sodium
tartrate not utilized.
Nitrate reduction: Negative.
Production of HS: Positive.
Antagonistic properties: Produces hy-
droxystreptomycin.
Type culture: IMRU 3557; ATCC 12,628.
109. Streptomyces griseochromogenes Yuku-
naga et al., 1955 (Fukunaga, K., Misato, T.
Ishu, I., and Asakawa, M. Bull. Agr. Chem
Soc. Japan 19: 181-188, 1955).
Morphology: Sporophores form closed spi-
rals on starch agar; there are no spirals, or
only curling tips, formed on sucrose nitrate
and glucose-asparagine agars. Spores spheri-
cal or oval, about 1.0 to 1.5 uw.
Sucrose nitrate agar: Growth spreading,
orange-cinnamon. Aerial mycelium white or
light neutral gray. No soluble pigment.
Glucose-asparagine agar: Growth — re-
stricted, ivory-yellow, penetrating into the
medium. No aerial mycelium, later white.
No soluble pigment.
Nutrient agar: Growth restricted, opales-
cent. No aerial mycelium. Soluble pigment
brown.
Potato: Growth
snuff-brown. Aerial mycelium
mouse-gray. Color of plug dark brown to
black around growth.
Gelatin: Growth wrinkled, yellowish in
wrinkled,
white to
abundant,
liquefied portion. Aerial mycelium white,
scant. Liquefaction medium. Soluble pig-
ment dark brown to black.
Milk: Growth as surface ring, brown. No
coagulation; peptonization begins in 8 days
at 37°C; not completed in 21 days.
Nitrate reduction: Positive.
Cellulose: No growth.
Tyrosine agar: Growth orange-colored. No
soluble pigment.
Invertase: Positive.
Carbon utilization: Glucose, p-fructose,
p-galactose, maltose, lactose, raffinose, pb-
mannitol, utilized.
inulin, p-sorbitol, dulcitol, salicin, sodium
DL-Inositol Rhamnose,
acetate, sodium citrate, sodium succinate not
utilized.
Antagonistic properties: Produces blasti-
cidins A, B, and C, active against fungi.
temarks: S. griseochromogenes belongs to
the group of chromogenic actinomycetes. S.
resistomycificus differs from S. griseochromo-
genes in the color of its aerial mycelium ob-
served on various media, and also in that it
produces an aerial mycelum on nutrient
agar and a soluble pigment in glucose-aspara-
gine agar. S. mirabilis has a different form
of aerial mycelium and a different optimum
temperature. S. flavochromogenes produces
an aerial mycelium on nutrient agar and a
grayish soluble pigment. S. olivochromogenes
assumes a dark brown or black color of
growth and shows an alkaline reaction in
milk medium. S. dzastatochromogenes is quite
similar to S. griseochromogenes in the ap-
pearance of its growth on several media, but
differs from it by producing a white or gray
aerial mycelium on nutrient agar, and also
by producing tyrosinase.
110. Streptomyces griseoflavus (srainsky,
1914) Waksman and Henrici, 1948 (Krain-
sky, A. Centr. Bakteriol. Parasitenk. Abt.
II, 41: 684, 1914).
Morphology: Sporophores straight, mono-
podially branched; no curvatures and no
spirals produced. Spores oblong, 1.0 to 1.2 u,
covered with short spines. (According to
Ettlnger et al. (1958), open, regular spirals
are formed.)
Sucrose nitrate agar:
brown to orange. Aerial mycelium gray to
Growth reddish-
yellowish-gray. Faint greenish-yellow soluble
pigment.
Glucose-asparagine agar: Growth citron-
yellow. Aerial mycelium powdery, greenish-
yellow changing to gray (Hoffmann, 1958).
Calcium malate agar: Growth yvellowish-
green-gray.
Nutrient Growth
covered with white to gray aerial mycelium.
agar: cream-colored,
Soluble pigment absent or, according to
Hoffmann (1958), greenish-gray.
Starch
brownish center. Aerial mycelium absent or
agar: Growth cream-colored with
powdery, gray. Hydrolysis limited.
THE ACTINOMYCETES, Vol. II
Potato: Growth lichenoid, yellowish. Ae-
rial mycelium powdery, white to gray. Mela-
nin-negative.
Gelatin: Growth cream-colored to brown-
ish, covered with white to yellowish-gray
aerial mycelium. Positive liquefaction. Mela-
nin-negative.
Milk: Growth cream-colored to yellowish;
aerial mycelium thin, white. No coagulation;
rapid peptonization.
Cellulose: Growth good. Greenish-yellow
soluble pigment, according to Hoffmann
(1958).
Nitrate: Strong reduction to nitrite.
Production of HS: Negative.
Invertase: Negative.
Antagonistic properties: According to
Waga (1953), a member of this group pro-
duced an antibiotic, griseoflavin; this anti-
biotic was later (Xuroya et al., 1958) found
to be identical with novobiocin. Another
antibiotic, grisamine, has also been reported.
Remarks: According to Jensen (1930), the
species is characterized by the grayish-yellow
color of its aerial mycelium, which never
assumes the distinct green shade of S. griseus.
A detailed study of the life cycle of this
organism has been made by Saito and Ikeda
(1958). They found that between the pri-
mary (vegetative phase) and the secondary
(sporulation phase) mycelium, there may be
‘
a transitional stage which comprises ‘‘nests,”’
“swollen bodies,” and ‘“‘clubs,’”’ correspond-
ing to the “initial cells’’ reported in the litera-
ture.
111. Streptomyces griseolus (Waksman,
1923) Waksman and Henrici, 1948 (Waks-
man, 8. A. Actinomyces 96. Soil Sci. 8: 121,
1919).
Morphology: Sporophores short, straight,
without spirals, some curling found on side
branches. Spores spherical to oval-shaped to
cylindrical.
Growth colorless,
thin, spreading, chiefly in the medium.
Aerial mycelium at first gray, later becoming
Sucrose nitrate
agar:
DESCRIPTION OF SPECIES OF STREPTOMYCES 223
pallid neutral gray, with yellowish tone.
Faint brownish soluble pigment.
Malate-glycerol agar: Growth brownish.
Aerial mycelium lght mouse-gray. Soluble
pigment faint brownish.
Glucose-asparagine agar: Growth cream-
colored, turning dark. Aerial mycelium deep
dull gray. No soluble pigment.
Nutrient agar: Brownish growth, with
smooth surface. Aerial mycelium white with
gray tinge. Soluble pigment brown. Mela-
nin-negative.
Potato: Growth cream-colored, becoming
black. Aerial mycelium white with greenish
tinge. Soluble pigment brown to black.
Gelatin: Yellowish flaky pellicle and sedi-
ment. Aerial mycelium white. Gradual lique-
faction. Faint browning of medium.
Milk: Growth abundant, pink pellicle.
Slow coagulation and good peptonization.
Starch media: Growth grayish-brown with
dark ring. Aerial mycelium gray. Slight hy-
drolysis of starch.
Cellulose: Scant growth.
Invertase: Negative.
Nitrate reduction: Positive.
Production of H.S: Negative.
Temperature: Optimum 25°C.
Antagonistic properties: Some strains
show considerable activity against various
bacteria. Several antibiotics (phagomycin,
fermicidin, anisomycin, oxytetracycline, gris-
eomycin) were isolated from cultures de-
scribed as strains of S. griseolus.
Habitat: Soil.
Remarks: Ettlinger et al. (1958) considered
this organism as related to S. olivaceus.
Krassilnikov (1949) considered it as a strain
of A. candidus. Hoffmann (1958) isolated a
strain of this species from potato scab. Al-
though this organism is usually described as
melanin-negative, Krassilnikov (1941) and
Hoffmann consider it as melanin-positive.
Type culture: IMRU 3325.
112. Streptomyces griseoluteus
i
1951 (Umezawa, H., Hayano, 5.,
Umezawa
et al.,
Maeda, K., Ogata, Y., and Okami, Y. J.
Antibiotics (Japan) 4: 34-40, 1951; Okami,
Y. zbid. 5: 477-480, 1952).
Morphology:
podial and irregular branching, flexible and
hooked. Spores oval to cylindrical, 1.0 to 1.2
by 1.8 to 2.2 yu.
Sucrose nitrate agar: Growth thin, color-
Sporophores with mono-
less to cream-colored. Margin plumose, pene-
trating into medium. Aerial mycelium pow-
dery, grayish-white to light drab. Soluble
pigment absent or yellowish-brown.
Glucose-asparagine agar: Growth § wrin-
kled, cream-colored. Aerial mycelium. thin,
white. Pigment reddish-brown.
Nutrient agar: Growth wrinkled, trans-
parent. Aerial mycelium thin, white, pow-
dery. Soluble pigment absent or yellowish-
brown.
Potato: Growth
cream-colored. Aerial mycelium dusty white,
thin. Plug becoming slightly brownish.
Gelatin: No growth.
Milk: Surface ring cream-colored. Aerial
abundant, wrinkled,
mycelium in form of white patches.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Production of H.S: Negative.
Antagonistic properties: Produces griseo-
lutein.
Type culture: IMRU 3674; 3729.
115. Streptomyces griseoplanus Backus et
al., 1957 (Backus, E. J., Tresner, H. D., and
Campbell, T. H. Antibiotics & Chemother-
apy 7: 532-541, 1957).
Morphology: Sporophores arise as tangled
and curved and often loosely spiraled chains
of spores. Spores globose to elliptical, 0.6 to
2 to 1.5 u (Fig. 40).
Growth
2 DY
Sucrose nitrate agar: colorless.
Aerial mycelium scant, white to gray.
Glucose-asparagine agar: Growth gray to
light pinkish. No aerial mycelium.
Nutrient agar: Growth ivory-yellow. No
aerial mycelium. Melanin-negative.
224 THE ACTINOMYCETES, Vol. II
FragureE 40. Sporophores of S. griseoplanus (Reproduced from: Backus, E. J.
Chemotherapy 7: 537, 1957).
Starch agar: Growth colorless to yellow-
ish. Aerial mycelium scant, white.
Potato plug: Growth light brownish. Ae-
rial mycelium scant, white to gray. Apex of
plug browned.
Milk: No growth.
Cellulose: No growth.
Production of HoS: Negative.
Carbon utilization: Extremely limited.
with ammonium sulfate as source of nitro-
gen. With aspartic acid, organism utilizes
L-arabinose, D-xylose, and glucose. Fair to
moderate growth on lactose, b-raffinose, pb-
trehalose, and salicin. Sucrose, p-fructose,
pb-mannitol, 7-inositol, adonitol, p-melezitose,
i.
et al. Antibiotics &
L-rhamnose, esculin,p-melibiose, and dextrin
utilized poorly or not at all.
Antagonistic properties: Produces anti-
biotic alazopeptin.
Habitat: Grassland soil.
femarks: This organism is closely related
to S. flavogriseus.
114. Streptomyces griseoruber Yamaguchi
and Saburi, 1955 (Yamaguchi, T. and Sa-
buri, Y. J. Gen. Appl. Microbiol. 1: 201-235,
1955).
Morphology: Aerial hyphae produce open
and closed spirals on sucrose nitrate and on
starch agars. Spores short, cylindrical, 0.5 to
0.9 by 0.9 to 1.2 p.
DESCRIPTION OF SPECIES OF STREPTOMYCES 225
nitrate agar: Growth reddish-
orange when freshly isolated, but changes to
Sucrose
colorless or whitish on repeated transfer.
Aerial mycelium powdery, drab-gray. No
soluble pigment.
Calcium malate agar:
glossy, pinkish-gray, later becoming dull
purplish. Aerial mycelium powdery, olive-
gray. Soluble pigment absent or faint yel-
Growth at first
lowish-brown.
Nutrient agar: Growth at first light olive-
gray, later becoming brown. Aerial myce-
lum absent or scanty, white. Soluble pig-
ment brown.
Starch agar: Growth light reddish-orange
to reddish-purple. Aerial mycelium powdery,
olive-gray. Soluble pigment faint yellow to
faint yvellowish-pink. Strong hydrolysis.
Potato: Growth wrinkled, at first olive-
gray to dark yellowish-brown, later becom-
ing dark reddish or black. Aerial mycelium
absent or scanty, white. Soluble pigment
deep purple to black.
Gelatin: Growth light yellowish-brown on
surface. No aerial mycelium. Soluble pig-
ment brown to lght yellowish-green. Lique-
faction weak to medium.
Milk: Growth deep brown at 37°C, but
cream-colored to light yellowish-brown, and
occasionally with pinkish tone at 25°C. Sol-
uble pigment grayish-yellow-brown at 37°C,
but sometimes faint yellowish-brown with
pinkish shade at 25°C. Coagulation begins in
3 days, followed by peptonization.
Cellulose: No growth.
Carbon utilization: Utilizes p-xylose, L-
arabinose, L-rhamnose, lactose, inositol, sali-
cin, sodium acetate; does not utilize sucrose,
raffinose, inulin, mannitol, sorbitol, citrate,
and succinate.
Antagonistic properties: Active
gram-positive and acid-fast bacteria; pos-
against
sesses antitrichomonal activity.
telated to the S.
and to S. erythrochromogenes.
temarks: ruber group
115. Streptomyces griseoviridis Anderson
et al., 1956 (Anderson, L. E., Ehrlich, J.,
Sun, 8. H., and Burkholder, P. R. Antibiot-
ics & Chemotherapy 6: 100-115, 1956).
Not S. griseoviridus.
Morphology: Sporophores straight or
curved, with open and closed spirals on
lateral branches. Spores spherical to ovoid,
0.6 to 1.5 by 0.8 to 2.1 yu.
Starch-ammonium sulfate agar: Growth
tan-gray to black. Aerial mycelium tan to
light brown.
Glycerol-asparagine agar: Growth light
vellow to gray. Aerial mycelium pink-tan to
eray-green.
Calcium malate agar: Growth yellow-tan
to gray. Aerial mycelium light brown. Slight
hydrolysis of starch.
Nutrient
ereen-gray to brown. Aerial mycelium light
agar: Growth yellow-tan to
gray-pink to light gray-green. Soluble pig-
ment light brown.
Glucose-tryptone light
brown to red-brown to dark brown or black.
agar: Growth
Aerial mycelium pink-gray to light brown
to green-brown. Soluble pigment brown; red-
brown near growth.
Gelatin: Fairly rapid liquefaction. Soluble
pigment light brown to dark brown.
Milk: Peptonization.
Starch: Hydrolysis.
Carbon utilization:
cellobiose, dextrin, galactose, glucose, glyc-
mannitol,
Utilizes arabinose,
erol, lactose, levulose, maltose,
mannose, rhamnose, starch, trehalose, and
xylose. Does not utilize esculin, adonitol,
dulcitol, z-inositol, inulin, melezitose, meli-
biose, raffinose, salicin, sorbitol, and sucrose.
Antagonistic properties: A source of gris-
eoviridin and viridogrisein (etamycin).
Habitat: Texas soil.
Type culture: IMRU 3735.
116. Streptomyces griseus Waksman and
Henrici, 1948 (Waksman and Henrici, Ber-
gey’s Manual, 6th ed. 1948, p. 948; Waks-
man, §. A. and Curtis, R. E. Soil Sei. 1:
119-120, 1916; Waksman, 8. A., Reilly,
DIG
Ei Cs"and Harris: DA. Je Bacteriol. 56:
259, 1948; Waksman, S. A. Proc. Natl.
Acad. Sei. U.S. 45: 1043-1047, 1959).
Synonyms: Actinomyces globisporus Wras-
silnikov, 1941. Actinomyces globisporus sub-
sp. streptomycinta (Waksman) Irassilnikov,
1949. Actinomyces streptomycini Wrassilni-
kov, 1957.
Morphology: Sporophores straight, pro-
duced in tufts (Fig. 41). Spores spherical to
oval, 0.8 by 0.8 to 1.7 uw; surface smooth (PI.
eben).
Sucrose nitrate agar: Growth thin, spread-
ing, colorless, becoming olive-buff. Aerial
mycelium thick, powdery, water-green. Pig-
ment insoluble.
Nutrient agar: Growth abundant, almost
transparent, cream-colored. Aerial mycelium
powdery, white to light gray. No soluble
pigment.
Glucose agar: Growth elevated in center,
radiate, cream-colored to orange, erose mar-
gin.
Substrate and aerial mycelium of
FIGURE 41.
S. griseus.
THE ACTINOMYCETES, Vol. II
Starch media: Growth thin, spreading,
transparent. Hydrolysis strong.
Tyrosine agar: Dark pigment often pro-
duced.
Potato: Growth wrinkled, yellowish to
brownish, covered with white, powdery aer-
ial mycelium.
Gelatin: Greenish-yellow or cream-colored
surface growth with brownish tinge. Rapid
liquefaction.
Milk: Cream-colored ring;
with rapid peptonization, becoming alkaline.
Cellulose: Scant to fair growth.
Nitrate reduction: Positive.
Pigments: Produces green or yellow solu-
ble pigment on calcium malate and succinate
media.
Production of HS: Negative.
Carbon sources: See S. griseinus.
Antagonistic properties: Strongly antago-
nistic. Produces antibiotic streptomycin, ac-
tive against a large number of bacteria and
actinomycetes, but not against most fungi or
viruses; also produces cycloheximide, active
upon fungi. Resistant to streptomycin-pro-
ducing organisms and to streptomycin.
Remarks: An extensive literature has ac-
cumulated on the nature of this organism
(KXoreniako and Nikitina, 1959), on its phage
sensitivity (xoerber et al., 1950), antibiotic
production (Waksman, 1949), ete.
Habitat: Soils, river muds,
chicken.
Type culture: IMRU 3463.
coagulation
throat of
117. Streptomyces hachijoensis Yamaguchi,
1954 (Yamaguchi, T. J. Antibiotics (Japan)
7A: 10-14, 1954).
Morphology: Aerial hyphae short, straight,
0.6 to 1.2 uw. Secondary verticils produced.
Spores cylindrical, 0.8 to 1.0 by 1.5 to 1.8 yu.
Sucrose nitrate agar: Growth restricted,
colorless; reverse yellowish. Aerial mycelium
white, changing to pinkish-buff. No soluble
pigment.
Calcium malate agar: Growth colorless to
DESCRIPTION OF SPECIES OF STREPTOMYCES
yellow. Aerial mycelium white to pale pink-
ish-buff. Soluble pigment absent.
Nutrient agar: Growth cream-colored,
wrinkled. Aerial mycelium powdery, shade of
pale ochraceous-buff. No soluble pigment.
Potato: Growth cream-colored to yellow-
ish, wrinkled, raised. Aerial mycelium white.
Soluble pigment around growth faint pur-
plish.
Blood agar: Growth yellow to brownish-
yellow. Aerial mycelium flocculent, white.
Soluble pigment dark. Positive hemolysis.
Gelatin: Growth yellow to brown. No
aerial mycelium. Rapid liquefaction.
Milk: Surface ring yellow to brown. Aerial
mycelium in form of white patches. Soluble
pigment pinkish to orange. Coagulation fol-
lowed by peptonization.
Nitrate reduction: Negative.
Cellulose: No growth.
Antagonistic properties: Produces an anti-
fungal agent, trichomycin, a member of the
candicidin group of antibiotics.
Remarks: Resembles S. rubrireticult. Bli-
nov (1958) described a variety (fuscatus) of
this species, as a producer of candicidin-
type antibiotics.
118. Streptomyces halstedii (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, S. A. and Curtis, R. E. Soil Sci.
J: 124, 1916; 8: 121, 1919).
Morphology: Sporophores
spirals. Spores oval or rod-shaped, 1.0. to
eZ 1.2 tOM Su:
Sucrose nitrate agar: Substrate growth
form closed
abundant, spreading, raised, at first light
colored, becoming dark to almost black.
Aerial mycelium white, turning dull gray.
No soluble pigment.
Glycerol malate agar: Growth dark. Aerial
mycelium deep mouse-gray.
Nutrient agar: Growth restricted, wrin-
kled, cream-colored. No aerial mycelium.
Melanin-negative.
Glucose-asparagine agar: Growth wrin-
227
kled, center elevated, edge lichenoid, color-
less, becoming brown. No aerial mycelium.
Potato: Growth abundant, moist, wrin-
kled, cream-colored with green tinge.
Gelatin: Small, cream-colored masses of
growth in bottom of tube. Rapid liquefae-
tion. No soluble pigment.
Milk:
and slow peptonization.
Starch media: Growth abundant, glossy,
brownish. No aerial mycelium. Rapid hy-
Cream-colored ring. Coagulation
drolysis.
Cellulose: No growth.
Nitrate: Reduction to nitrite.
Production of H»S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Strongly antag-
onistic; some strains show only antifungal
activity; some strains produce carbomycin.
Habitat: Soil.
Remarks: Several closely related forms
have been described. According to Ettlinger
et al. (1958), the strains examined produce
no spirals and belong to S. oldvaceus. Ac-
cording to Okami and Suzuki (1958), the
sporophores are wavy, seldom forming hooks
or primitive spirals. Gause et al. (1957) de-
scribed a closely related form as A. griseoin-
carnatus.
Type culture: IMRU 3328.
119. Streptomyces hawaiiensis Cron et al.,
1956 (Cron, M. J., Whitehead, D. I°., Hooper,
I. R., Heinemann, B., and Lein, J. Anti-
biotics & Chemotherapy 6: 63-67, 1956).
Morphology: Sporophores produce spirals
on some media. Spores oval, 0.6 to 0.8 by
O57 to des we
Sucrose nitrate agar: Growth faint yellow.
Aerial mycelium sparse, white to flesh-col-
ored. Soluble pigment faint tan or absent.
Glucose-asparagine agar: Growth light
brownish. Aerial mycelium moderate, white
to gray. Soluble pigment faint tan or absent.
Nutrient agar: Growth gray with light
Aerial mycelium limited,
brown reverse.
223
wood-ash to steel-gray. Soluble pigment
brown.
Potato: Growth gray. Aerial mycelium
limited, gray. Soluble pigment dark brown,
almost black.
Milk: No coagulation or peptonization.
Slight acid reaction. Soluble pigment green-
ish-brown.
Gelatin: Slight liquefaction at 26°C in 19
days. Soluble pigment brown.
Starch: Shght hydrolysis in 7 days at 28-
30°C.
Blood agar: No hemolysis. Soluble pig-
ment black.
Production of H.S: Positive.
Carbon Utilizes
rhamnose, glucose, galactose, fructose, su-
utilization: arabinose,
crose, maltose, lactose, xylose, raffinose, cell-
obiose, dextrin, inulin, soluble starch, glyc-
erol, inositol, mannitol, sodium acetate,
sodium citrate, and calcium malate. Does
not utilize sorbitol, dulcitol, sodium oxalate,
sodium salicylate, sodium tartrate, and so-
dium succinate.
Antagonistic properties: Produces a poly-
peptide antibiotic, bryamycin.
Remarks: S. hawaziensis is a chromogenic
form which produces a soluble, dark brown
pigment on protein media and a white to
eray aerial mycelium. The organism is fur-
ther characterized by spiral formation in the
aerial mycelium and weak proteolytic ac-
tivity in gelatin and milk.
Streptomyces phaeofaciens possesses cul-
tural and morphological characteristics simi-
lar to those of S. hawaziensis, but differs in
its rapid peptonization of milk and produc-
tion of an antifungal substance inactive on
bacteria.
S. hawariensis resembles S. aureus in that
spiral formation occurs with both cultures
and both form soluble brown pigments in
organic media. They differ in that S. aureus
liquefies gelatin to a greater extent. S. ha-
wariensis 1s also similar to S. bekiniensts in
some of its cultural properties; both produce
THE ACTINOMYCETES, Vol. II
a white aerial mycelium which becomes
gray-colored; the sporulation of S. hawazien-
sis takes place in the form of spirals in its
aerial mycelium, whereas S. bekiniensis is
completely devoid of spirals and produces an
alkaline reaction accompanied by hydroly-
sis in milk.
Type culture: ATCC 12,236.
120. Streptomyces hiroshimensis Shinobu,
1955 (Shinobu, R. Seibutsugakkaishi 6: 43—
46, 1955).
Morphology: Sporophores produce verti-
cils of the Nitella type, both primary and
secondary. No spirals. Spores elliptical to
oval,0:8 to 1.24 (Pl. V¥,-Gb).
Sucrose nitrate agar: Growth poor, re-
stricted, pink. Aerial mycelium scant, pale
pink to pinkish-white.
Calcium malate agar: Growth slow. Aerial
mycelum pale cinnamon-pink. Soluble pig-
ment brownish.
Glucose-asparagine agar: Growth good,
reddish-pink. Aerial mycelium pink to pur-
plish-pink. Soluble pigment usually absent,
sometimes pale brown.
Nutrient agar: Growth
Aerial mycelium absent, or scant, pale pink
to pinkish-white. Soluble pigment brownish-
reddish-brown.
orange.
Starch agar: Growth red to purplish-red.
Aerial mycelium pink to pale pink.
Potato plug: Growth deep pinkish-red to
Aerial mycelium
pinkish-white. Soluble pigment brownish-
black.
Gelatin: Growth pale reddish-brown. Aer-
pinkish-
brownish-black. scant,
ial mycelium absent, or scant,
white. Soluble pigment pale reddish-brown.
Rapid liquefaction.
Milk: Growth deep pinkish-red. Aerial
mycelium pinkish-white to pink. Soluble pig-
ment brown with reddish tinge. No coagu-
lation; rapid peptonization.
Nitrate reduction: Strong.
Starch: Rapid hydrolysis.
Tyrosinase reaction: Positive.
DESCRIPTION OF SPECIES OF STREPTOMYCES 229
Cellulose: Not attacked.
Carbon utilization: I'ructose and inositol
well utilized; xylose, rhamnose, sucrose, lac-
tose, raffinose, and mannitol not utilized;
galactose and trehalose slightly utilized.
Antagonistic properties: Inhibits growth
of gram-positive bacteria and fungi.
Source: Isolated from soil in Hiroshima,
Japan.
temarks: Resembles S. rubrireticuld.
121. Streptomyces hirsutus Ettlinger et al.,
1958 (Ettlinger, L., Corbaz, R., and Hiitter,
R. Arch. Mikrobiol. 31: 344, 1958).
Morphology: Sporophores
podially branched; short open spirals with
about three coils are produced. Spores cov-
long, mono-
ered with narrow, long spines (PI. II, k).
Glycerol nitrate agar: Growth colorless.
Aerial mycelium at first milky white, later
leek-green.
Glucose-asparagine agar: Growth color-
less. No aerial mycelium.
Glycerol malate agar: Growth at
milky white; later covered with aerial my-
celium gradually colored leek-green.
Starch-KNOs; agar: Growth milky white.
Aerial mycelium leek-green. Starch hydro-
first
lyzed.
Gelatin: Growth whitish-yellow, covered
with light green aerial mycelium. Slow lique-
faction. No soluble pigment. Melanin-nega-
tive.
Potato: Growth colorless. Aerial mycelium
at first white, later leek-green.
Milk: Pellicle heavy, light yellow. Aerial
mycelium white-gray. Rapid coagulation; no
peptonization.
Antagonistic properties: None.
Habitat: Soil in Switzerland.
temarks: Some of the cultures described
by Gause et al. (1957), such as A. acrimycini
and A. acrimycint var. globosus, are closely
related to this organism.
(Bostroem
Waksman
122. Streptomyces hominis
1890; emend. Waksman, 1919)
and Henrici, 1948. (Bostroem, E. Beitr.
pathol. Anat. allgem. Pathol. 9: 1-240,
1890; Waksman, S. A. Soil Sci. 8: 129-130,
1919.
Synonyms: Streptothrix hominis Fouler-
ton, 1899. Oospora hominis Ridet, 1911.
Morphology: Sporophores straight. A few
dextrorse spirals on glycerol synthetic me-
dia.
Sucrose nitrate agar: Growth white with
shade of yellow, turning brownish with age.
Aerial mycelium white with olive tinge. No
soluble pigment.
Glycerol malate agar: Growth yellowish.
Aerial mycelium with olive-green tinge.
Nutrient agar: Growth yellowish. Aerial
mycelium white. No soluble pigment.
Starch: Hydrolysis good.
Potato: Growth yellowish to orange, be-
coming brown. Aerial mycelium white. Color
of plug unchanged, later becoming brown.
Gelatin: Growth cream-colored. No aerial
mycelium. No soluble pigment.
Milk: Rapid coagulation and peptoniza-
tion.
Nitrate reduction: Positive.
Sucrose: Not inverted.
Production of HoS: Negative.
Habitat: Supposed to have been isolated
from abscess of palm of hand; probably an
ar contamination. Appears to be related to
the S. griseus series.
125. Streptomyces humidus Nakazawa and
Shibata, 1956 (Belgian Patent 533,386. Ta-
keda Pharmaceutical Industries Ltd., Ja-
pan, March 24, 1956; Proc. Japan Acad.
32: 648-653, 1956).
Morphology: Sporophores form spirals.
PANTO Te
Growth
Spores oval, 1 to 1.5 by
nitrate agar:
Aerial mycelium white. No soluble pigment.
Nutrient agar: Growth colorless. No aer-
ial mycelium. No soluble pigment.
Sucrose colorless.
Glucose-asparagine agar: Growth color-
less. Aerial mycelium white to smoke-gray
or vinaceous-chamois. No soluble pigment.
230
Calcium malate agar: Growth colorless,
becoming yellowish. Aerial mycelium white.
No soluble pigment.
Starch agar: Growth colorless to cream-
colored. Aerial mycelium white to pale
smoke-gray. No soluble pigment.
Potato: Growth colorless. Aerial mycelium
white to smoke-gray; black, moist speckles.
No soluble pigment.
Gelatin: Growth colorless. No aerial my-
celium. No soluble pigment. Moderate lique-
faction.
Milk: Growth colorless. Aerial mycelium
white. No soluble pigment. Slow peptoniza-
tion.
Nitrate reduction: Positive.
Carbon utilization: b-xylose, L-arabinose,
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Fraure 42. 8S. hygroscopicus (Reproduced from:
Tresner, H. D. and Backus, EH. J. Appl. Micro-
biol. 4: 246, 1956).
THE ACTINOMYCETES, Vol. II
L-rhamnose, b-fructose, galactose, maltose,
lactose, D-mannitol, salicin utilized. Sucrose,
D-raffinose, inulin, p-sorbitol, dulcitol, 2-ino-
sitol, sodium acetate, sodium citrate not uti-
lized.
Antagonistic properties: Produces an anti-
biotic, acidomycin, said to be dihydrostrep-
tomycin (see also Imamura et al., 1956).
Remarks: S. huwmidus is closely related to
S. hygroscopicus; growth of the latter on
agar media and on potato is cream-colored
to yellow to brown.
124. Streptomyces hygroscopicus (Jensen,
1951) Waksman and Henrici, 1948 (Jensen,
H. L. Proc. Linnean Soc. N. 8. Wales 56:
30/—358, 1931).
Morphology: Sporophores monopodially
branched, with narrow compact, sinistrorse
spirals, situated as dense clusters on the
main stems of the sporophores (Fig. 42).
Spores oval, 0.8 to 1.0 by 1:0 to 1:22,
smooth (PI. II nf, Pl. IV Gb).
nitrate agar: Growth folded,
white to cream-colored, later sulfur-yellow
to yellowish-gray, with golden to light orange
reverse. Aerial mycelium scant, white to ash-
gray. Soluble pigment golden to light orange.
Glucose-asparagine agar: Growth cream-
colored to straw-yellow, later dull chrome-
vellow to brownish-orange. Aerial mycelium
Sucrose
dusty white to pale yellowish-gray; later
small, moist, dark violet-gray to brownish
patches produced, gradually spreading over
the whole surface. Soluble pigment light
yellow.
Nutrient agar: Growth wrinkled, cream-
colored, later vellowish-gray with yellowish-
brown reverse. Aerial mycelium scant, white.
Potato: Growth raised, wrinkled, cream-
colored, later yellowish-gray to dull brown-
ish. Aerial mycelium absent or trace of
white. Melanin-negative.
Gelatin: Liquefaction slow. No soluble
pigment.
Milk: No coagulation; positive peptoniza-
DESCRIPTION OF SPECIES OF STREPTOMYCES 231
tion. The reaction becomes faintly acid (pH
6.0 or less).
Starch: Hydrolysis.
Cellulose: Ready decomposition by some
strains.
Nitrate reduction: None with sucrose as
source of energy.
Sucrose inversion: Positive.
Production of H.S: Negative.
Antagonistic properties: Produces hygro-
mycin, an antibiotic active against mycobac-
teria and roundworms (see also Pagano ef al.,
1953).
Habitat: Soil.
Remarks: Tresner and Backus (1956)
made a comprehensive study of 18 cultures
representing S. hygroscopicus and closely re-
lated forms. They came to the conclusion
that the following three properties are the
fundamental the organ-
ism: (1) sporophores terminate in tight spi-
rals of a few to many turns, plus a clustering
of such sporophores along hyphae; (2)
brownish-gray (mouse-gray to benzo-brown)
characteristics of
LD)
spore color on favorable media; (3) distine-
tive hygroscopic character on some agar
media. The characteristic feature, not
equally distinct in all strains, however, is the
fact that the aerial mycelium on synthetic
media becomes moistened and exhibits dark,
glistening patches; when touched with a
needle, these patches prove to be moist,
smeary masses of spores. Shape and size of
spores, color of substrate growth, formation
of soluble pigments, growth on potato,
growth on milk, cellulose decomposition, and
carbon and nitrogen utilization were con-
sidered by Tresner and Backus as variable
properties. They considered S. platensis and
S. endus as closely related.
Other related forms, such as A. kurssano-
vi and A. nigrescens, have been described by
Gause et al. (1957). Ettlinger et al. (1958)
also included S. platensis and S. rutgersensis
var. castelarense in this group. The relation-
ship of this species to S. violaceoniger has
been indicated by Nomi (1960b). Vavra et al.
(1959) described a variety decoyicus that
differed in certain minor properties. A num-
ber of other varieties of this organism have
been described, such as odoratus (Yiintsen
et al., 1956) and angustmyceticus (Takahashi
and Amano, 1954).
125. Streptomyces intermedius (Kriiger-
emend. Wollenweber, 1922) Waksman (Wol,
lenweber, H. Ber. deut. botan. Ges. 39: 26,
1922).
Morphology: Sporophores straight, wavy,
frequently arranged in fascicles or clumps
No spirals. Spores round to oval; 0.9 to 1.0
by 0.7 uy.
Glycerol nitrate agar: Substrate growth
cream-colored to brown; sometimes dark
green to greenish-brown. Aerial mycelium
thin, gray to dark gray.
Glucose-asparagine agar: Growth brown-
ish with greenish shade. Aerial mycelium
dark gray. No soluble pigment.
Nutrient agar: Growth much __ folded,
cream-colored. Aerial mycelium white. Sol-
uble pigment faintly golden, occasionally
green to olive-green; on continued cultiva-
tion, green color tends to become cream-
colored to brownish.
Glucose-peptone Growth good,
brownish. Aerial mycelium heavy, cream-
colored to dark gray. No soluble pigment.
Potato: Growth folded, brown to greenish-
agar:
brown. Aerial mycelium dark gray. Soluble
pigment olive-green. Melanin-negative.
Gelatin: Growth thin, colorless to faintly
brown, dropping to bottom. Slow liquefac-
tion. Greenish-brown pigment.
Milk: Surface growth heavy, cream-col-
ored. No aerial mycelium. No coagulation,
good peptonization.
Starch: Good hydrolysis.
Cellulose: Growth good, olive-green. Aer-
ial mycelium dark gray.
Sucrose: Inversion slow.
Nitrate reduction: Limited.
Habitat: Potato scab.
~)
bo
Remarks: Above description was supple-
mented by Hoffmann (1958). Krassilnikov
(1949) considers this organism as a variety of
A. cretaceus.
Type culture: IMRU 3329.
126. Streptomyces tpomoeae (Person and
Martin, 1940) Waksman and Henrici, 1948
(Person, L. H. and Martin, W. J. Phyto-
pathology 30: 913, 1940).
Morphology: Spores oval to elliptical, 0.9
to 1.3 by 1.8 to 1.8 yw.
Sucrose nitrate agar: Growth abundant,
wrinkled, olive-yellow. No aerial mycelium.
Nutrient agar: Growth moderate, in form
of small, shiny colonies, both on the surface
and imbedded in the medium, silver-colored.
Starch agar: Growth moderate, smooth,
ivory-colored. Aerial mycelium white with
patches of bluish-green. No soluble pigment.
Complete hydrolysis after 12 days.
Potato: Growth moderate, shiny, wrin-
kled, light brown. No aerial mycelium. No
soluble pigment.
Gelatin: Growth scanty, after 25 days at
20°C. No aerial mycelium. No soluble pig-
ment. Some liquefaction.
Milk: Ring on surface. No visible coagu-
lation; positive peptonization.
Cellulose: No growth.
Nitrate reduction: Positive.
Antagonistic properties: Positive.
Habitat: Lesions caused by sweet-potato
disease.
Type culture: IMRU 3476.
kanamyceticus Okami
and Umezawa, 1957 (Umezawa, H., Ueda,
M., Maeda, K., Yagishita, K., Kondo, S.,
Okami, Y., Utahara, R., Osato, Y., Nitta,
KX., and Takeuchi, T. J. Antibiotics (Japan)
LOA: 181-188, 1957).
127. Streptomyces
Morphology: Sporophores flexible and
hooked, no true spirals.
Glycerol nitrate agar: Growth at first
colorless, later lemon-yellow. Aerial myce-
lium white to yellow, occasionally with a
THE ACTINOMYCETES, Vol. II
greenish or faint pinkish tinge. Soluble pig-
ment occasionally produced, faint brown.
Growth color-
less to yellow with faint pinkish-white, and
yellow or hay-colored reverse. Aerial myce-
hum scant; develops from center of colony.
white to faint pinkish-white to greenish-
Glucose-asparagine agar:
yellow or yellow. Soluble pigment occasion-
ally produced, faint brown.
Calcium malate agar: Growth yellow.
Aerial mycelium white-yellow.
Nutrient agar: Growth cream-colored.
Aerial mycelium absent or white. No soluble
pigment.
Potato: Growth wrinkled, faint yellowish-
brown to yellow. Aerial mycelium scant,
white. No soluble pigment. Substrate be-
neath growth occasionally changes to brown.
Gelatin: Liquefaction positive. No soluble
pigment. Melanin-negative.
Milk: Growth colorless. Aerial mycelium
absent or white. Coagulation and peptoni-
zation doubtful.
Blood agar: Growth wrinkled, grayish-
reddish-brown. No aerial mycelium. No sol-
uble pigment.
Starch: Hydrolyzed.
Carbon utilization:
dextrin, fructose, galactose, glycerol, mal-
Utilizes arabinose,
tose, mannitol, mannose, raffinose, starch,
sucrose, and succinate. Does not utilize ino-
sitol, inulin, lactose, rhamnose, sorbose, xy-
lose, and acetate. Some strains grow slightly
on eseulin, salicin, sorbitol, and citrate.
Antagonistic properties: Produces kana-
mycin, an antibiotic related to the neomycin
group.
temarks: Closely related to S. albido-
flavus, S. liesket, and S. alboflavus. Okami
et al. (1959a) made a detailed study of the
mutants produced by this organism.
128. Streptomyces kentuckensis Barr and
Carman, 1956 (Barr, IF. S., and Carman,
P. I. Antibiotics & Chemotherapy 6: 286—
289, 1956).
Morphology: thick-
Aerial mycelium
DESCRIPTION OF SPECIES OF STREPTOMYCES 233
walled, generally not branched; sporophores
straight; do not form spirals. Spores oblong
to oval, 0.5 by 0.5 to 1.5 uw. Spores produced
by fragmentation of the hyphae in substrate
mycelium are generally smaller than those
formed from aerial hyphae.
Nutrient agar: Growth gray to yellow. No
soluble pigment. Melanin-negative.
Gelatin: Slow liquefaction. No soluble pig-
ment.
Potato: Mycehum gray. Plug darkened.
Milk: Peptonization positive.
Nitrate reduction: Positive.
Production of H.S: Negative.
Carbon utilization: Readily utilizes vari-
ous pentoses, hexoses, disaccharides, ace-
tate and citrate; slight utilization of rham-
nose, inulin, glycerol, inositol, mannitol; does
not utilize dextran or salts of oxalic, succinic,
and salicylic acids.
Antagonistic properties: Effective against
gram-positive and some gram-negative bac-
teria. Produces antibiotic raisnomycin.
Remarks: Pridham et al. (1958) consider
this organism as a member of the bzverticilla-
tus group.
Type culture: ATCC 12,691.
129. Streptomyces kimbert (Erikson, 1935)
Waksman (Erikson, D. Med. Research
Council (Brit.) Spec. Rept. Ser. No. 203:
14-15, 1935).
Morphology: Growth made up of long,
straight, profusely branching filaments. Ae-
rial mycelium produces short and straight
sporophores. Spores small, round.
Sucrose nitrate agar: Growth
cream-colored. Aerial mycelium powdery,
white.
Glucose-asparagine agar: Growth cream-
colored. Aerial mycelium white.
Nutrient agar: Growth moist, cream-col-
ored. Aerial mycelium powdery, white.
Gelatin: Colonies smooth, shining, float-
ing on liquefied medium. Aerial mycelium
powdery, white. Good liquefaction. No sol-
uble pigment.
moist,
Milk: Coagulation, slow peptonization.
Surface ring pinkish-brown; medium later
becomes dark brown.
Starch: No hydrolysis.
Source: Blood culture of a woman with
acholuric jaundice. No record concerning
actual pathogenicity.
130. Streptomyces kitasatoensis Hata et al.,
1953 (Hata, T., Koga, F., and Kanamori,
H. J. Antibiotics (Japan) 6A: 109-112,
1953).
Morphology: Sporophores produce pri-
mary and secondary verticils. A few spirals
were also found. Spores oval or cylindrical,
1.9 to 1.3 by 0.9 u.
Sucrose nitrate agar: Growth yellow to
light vellowish-brown. Some strains form no
aerial mycelium even after prolonged culti-
vation; others form thick grayish-white ae-
rial mycelium. Soluble pigment light yellow-
ish.
Glucose-asparagine agar: Growth brown
to dark brown, restricted, with raised center.
Aerial mycelium thin, grayish or mouse-
grayish. Soluble pigment brown.
Nutrient agar: Growth brown, restricted,
with
Soluble pigment brown.
Starch Growth
vellow to yellowish-brown. Aerial mycelium
raised center. No aerial mycelium.
agar: colorless; reverse
yvellowish-white, cottony or flocculent.
Tyrosine agar: Growth brown to dark
brown. Aerial mycelium grayish-white, thin,
later becoming cottony. Tyrosinase reaction
positive.
Potato: and
wrinkled. No aerial mycelium. Color of plug
Growth yellowish-brown
light brown.
Gelatin: Growth dark brown. Soluble pig-
ment dark brown. Liquefaction slow at be-
ginning, but complete liquefaction 4 weeks
later.
Milk: No coagulation; slow peptonization.
No clearing of milk, but heavy brown pre-
cipitate on bottom; color of liquid in upper
portion brownish.
{ THE
)
Starch: Hydrolysis positive.
Cellulose: No decomposition.
Nitrate reduction: Positive.
Production of H.S: Positive.
Carbon utilization: Utilizes
starch, dextrin, glycerol, galactose, maltose,
glucose,
sucrose, trehalose, inositol, sorbitol, sodium
succinate, sodium citrate, and sodium ace-
tate. Does not utilize xylose, raffinose, rham-
nose, lactose, arabinose, mannose, mannitol,
inulin, dulcitol, fructose, salicin, or esculin.
Antagonistic properties: Produces an anti-
biotic, leacomycin.
temarks: S. kitasatoensis is similar to S.
reticuli in morphology of the mycelium, cul-
tural characteristics, and utilization of car-
bon sources, but different in several other
respects.
131. Streptomyces kitasawaensis Harada
and Tanaka, 1956 (Harada, Y. and Tanaka,
S. J. Antibiotics (Japan) 9A: 113-117, 1956).
Morphology: Sporophores straight ; no spi-
rals.
Sucrose nitrate agar: Growth cream to
yellow. Aerial mycelium white with pale
pinkish tinge.
Calcium malate agar: Growth cream-col-
ored. Aerial mycelium white. No soluble
pigment.
Glucose-asparagine agar:
brownish tinge. Aerial mycelium white with
Growth has
grayish tinge. Soluble pigment pale yellow-
ish-brown to pale greenish-yellow.
Nutrient agar: Growth brownish. Aerial
mycelium absent or scarce. Soluble pigment
dark brown.
Starch agar: Growth pale grayish-brown
to pale blackish-brown. Aerial mycelium
white. Soluble pigment pale greenish-yellow
to pale yellowish-brown.
Gelatin: Growth white to gray. Aerial my-
celiuum absent or scarce. Soluble pigment
dark brown. No or weak liquefaction.
Potato: Growth brown. Aerial mycelium
white. Soluble pigment dark brown.
ACTINOMYCETES, Vol. II
Milk: Growth in form of dark brownish
ring. Coagulation and peptonization.
Carbon utilization: Utilizes p(+)xylose,
D-mannitol, L-arabinose, salicin. Does not
utilize L(-++-)rhamnose, b-maltose.
Antagonistic properties: Produces an anti-
tumor substance, carzinocidin.
Habitat: Soil.
1959
203,
132. Streptomyces lanatus Frommer,
(Frommer, W. Arch. Mikrobiol. 32:
1959).
Morphology: Sporophores long, straight
or wavy, with short side branches; the ends
of these are more entangled than spiral-
shaped.
Glycerol-sucrose agar: Growth abundant,
cottony, with red-brown reverse. Soluble
pigment brown to dark red-brown.
Glucose-asparagine agar: Growth
brown. Aerial mycelium velvety to cottony,
rose to gray-green. Soluble pigment brown-
rose-
ish.
Calcium malate agar: Growth colorless to
yellowish. Aerial mycelium powdery, gray to
eray-green. No soluble pigment.
Nutrient agar: Growth yellow-brown. Ae-
rial mycelium cream-colored or lacking. Sol-
uble pigment yellow-brown to dark brown.
Starch: Weak hydrolysis.
Potato: Growth yellow-brown. Aerial my-
celium powdery, white. Soluble pigment
black.
Gelatin: Growth yellow. Aerial mycelium
yellow. Soluble pigment brown to red-brown.
Liquefaction medium.
Milk: Growth abundant, dark
Aerial mycelium powdery, cream-colored.
Coagulation, slow peptonization.
Cellulose: No growth.
Antagonistic properties: Produces actino-
brown.
mycin.
femarks: Closely related to S. purpureo-
chromogenes and S. phaeochromogenes.
133. Streptomyces lavendulae (Waksman
and Curtis, 1916) Waksman and Henrici,
DESCRIPTION OF SPECIES OF STREPTOMYCES
1948 (Waksman, S. A. and Curtis, R. E.
Soil Sci. 1: 126, 1916; 8: 130, 1919).
Morphology: Sporophores long, mono-
podially branched; short, compact spirals of
the dextrorse type, 5 to 8 uw in diameter;
spirals sometimes open. Some strains form
no spirals, Okami (1956).
Spores! oval 1:0'sto 2" by “k6to 2:0
smooth (Pl. V, Ea).
Sucrose nitrate agar: Growth thin, spread-
ing, colorless to cream-colored. Aerial my-
according to
celium cottony, white, becoming vinaceous-
lavender. No soluble pigment.
Glycerol malate agar: Growth cream-col-
ored. Aerial mycelium lavender. No soluble
pigment.
Glucose-asparagine agar: Growth yellow-
ish. Aerial mycelium white with lavender
tinge. No soluble pigment.
Nutrient agar: Growth wrinkled, gray. No
aerial mycelium. Soluble pigment brown.
Starch agar: Growth restricted, glistening,
transparent, rose-colored. Aerial mycelium
lavender. Good hydrolysis of starch.
Potato: Growth thin, wrinkled, cream to
yellowish. No aerial mycelium. Soluble pig-
ment black.
Gelatin: Surface growth creamy to brown-
ish. Aerial mycelium absent or white. Lique-
faction slow. Soluble pigment brown.
Milk: Cream-colored ring. No coagulation;
good peptonization.
Cellulose: Growth scant.
Nitrate reduction: Positive.
Production of H.S: Positive.
Temperature: Optimum 37°C.
Antagonistic properties: Various strains
of this organism produce antibiotics. One
such antibiotic, streptothricin, is active
both in vitro and in vivo against various
gram-positive and gram-negative bacteria,
fungi, and actinomycetes. Certain other
strains produce an antiviral agent, ehrlichin.
Habitat: Soil.
Remarks: Numerous strains and varieties
of this organism have been isolated. It is
235
sufficient to mention S. lavendulae var.
japonicus, and several of the cultures listed
by Gause et al. (1957), notably A. flavotri-
cint, A. toxytricini, and A.
linger et al. (1958) considered S. acidomyceti-
violascens. Ett-
cus and S. virginiae as members of this
group. KXrassilnikoy (1949) considered this
species as a variety of A. chromogenes.
Okami (1956) and Rangaswami (1958) made
a detailed study of numerous representatives
of this species or species-group.
Morais et al. (1958) described a variety of
S. lavendulae as brasilicus, the aerial my-
celium being red-pink or red-brown but not
lavender and not utilizing salicin.
Type species: IMRU 3440.
154. Streptomyces lieskei (Duché, 1934)
Waksman and Henrici, 1948 (Duché, J. Les
actinomyces du groupe albus. P. Lechevalier,
Paris, 1934).
Morphology: Sporophores form oval
spores.
Glucose-asparagine agar: Growth cream-
colored, later yellowish to green. Aerial my-
celium white, later yellowish, growing from
the edge toward the center. Soluble pigment
dirty yellow to yellow-green.
Nutrient agar: Growth cream-colored. Ae-
rial mycelium white. Soluble pigment yellow-
ish.
Gelatin: Aerial
mycelium white. No soluble pigment. Liq-
Growth cream-colored.
uefaction rapid.
Milk: Growth No aerial
mycelium. Peptonization without coagula-
tion. After 20 days the whole milk becomes
a clear yellowish liquid.
Tyrosine medium: Growth rapid. Aerial
mycelium whitish-yellow. Soluble pigment
cream-colored.
yellowish to orange-yellow.
Coagulated serum: Growth colorless. Liq-
uefaction rapid.
temarks: Related to S. alboflavus and S.
albidoflavus.
135. Streptomyces limosus Lindenbein,
236 THE ACTINOMYCETES, Vol. II
1952 (Lindenbein, W. Arch. Mikrobiol. 17:
361-383, 1952).
Morphology: Substrate mycelium divides
completely into coccoidal pieces. Some simi-
larity to Nocardia. Aerial mycelium gray,
produced in certain media.
Glycerol nitrate agar: Growth colorless,
later becoming deep yellow. No aerial my-
celium. Soluble pigment citron-yellow.
Glucose-asparagine agar: Growth lemon-
yellow, later becoming black with yellow
reverse. Aerial mycelium white, later ash-
eray. Soluble pigment lemon-yellow.
Glycerol malate agar: Growth dark yel-
low. Aerial mycelium white, later ash-gray.
Soluble pigment golden yellow.
Nutrient agar: Growth lght brown. No
aerial Soluble light
brown. Melanin-positive.
mycelium. pigment
Glucose-peptone Growth yellow-
brown. Aerial mycelium ash-gray. Soluble
agar:
pigment yellow-brown.
Starch-nitrate agar: Growth
vellow. Aerial mycelium gray-white. Soluble
brownish-
pigment light yellow. Hydrolysis strong.
Potato: Growth brownish-yellow. Aerial
mycelium Soluble
lemon-yellow to sulfur-vellow.
Gelatin: Growth yellow-brown. No aerial
eray-white. pigment
mycelium. Soluble pigment dark brown.
Liquefaction complete.
Milk: Growth lichenoid, light
Aerial mycelium gray-white. Soluble pig-
vellow.
ment hght brown. Strong peptonization.
Cellulose: No growth.
Antagonistic properties: None.
Source: Isolated from the slime of a river
bank.
temarks: Related to S. flavovirens.
136. Streptomyces ipmani (Waksman and
Curtis, 1916) Waksman and Henrici, 1948
(Waksman, 8. A. and Curtis, R. E. Soil
Sci. 1: 123, 1916: 8: 121, 1919).
Morphology: Sporophores straight, no spi-
rals. Spores oval, 0.8 to 1.1 by 1.0 to 1.5 wu.
Sucrose nitrate agar: Growth abundant,
raised, colorless, becoming light brown and
wrinkled. Aerial mycelium white, turning
gray to dark gray. No soluble pigment.
Glycerol malate agar: Growth colorless,
dark brown. Aerial
mouse-gray. No soluble pigment.
becoming mycelium
Glucose-asparagine agar: Growth spread-
ing, light yellow. No aerial mycelium. No
soluble pigment.
Nutrient agar: Growth wrinkled, glossy,
vellow. No aerial mycelium. No soluble pig-
ment.
Potato: Growth abundant, wrinkled,
cream-colored. Aerial mycelium white to
gray. Soluble pigment purplish.
Gelatin: Cream-colored, flaky sediment.
Aerial mycelium white-gray. Liquefaction
medium to rapid. Melanin-negative.
Milk: Cream-colored
and peptonization.
Starch media: Growth transparent, be-
ring. Coagulation
coming dark with age. No aerial mycelium.
Hydrolysis medium.
Cellulose: No or very scant growth.
Invertase: Positive.
Nitrate reduction: Positive.
Production of HoS: Negative.
Temperature: Optimum 25°C.
Antagonistic properties: Good, though
some strains show no activity.
Habitat: Soil.
femarks: Ettlinger e¢ al. (1958) consider
this organism as a strain of S. griseus.
reported that their
strain produced a grayish-yellow-buff aerial
Tresner and Danga
mycelium. Krassilnikov (1949) considered it
as a variety of A. viridis.
Type culture: IMRU 3331.
137. Streptomyces loidensis (Millard and
Burr, 1926) Waksman (Millard, W. A., and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores. straight and
spiral-forming. Spores cylindrical to spheri-
eal, 0.9 to 1.0 by 0.9 to 0.95 wu.
DESCRIPTION OF SPECIES OF STREPTOMYCES 237
Sucrose nitrate agar: Growth thin, flat,
gray to yellowish-olive. Aerial mycelium
scant, olive-colored. Soluble pigment yellow.
Nutrient potato agar: Growth good, gray.
Aerial mycelium olive-buff. Soluble pigment
golden brown.
Gelatin: Growth gray. Aerial mycelium
scant, white. Liquefaction rapid. Soluble
pigment yellow.
Milk: Surface growth excellent. Aerial
mycelium white. Coagulation and rapid pep-
tonization.
Starch: Hydrolysis.
Nitrate reduction: None.
Temperature: Grows well at 37.5°C.
Habitat: Potato scab.
138. Streptomyces longisporoflavus (xras-
siinikov, 1941) Waksman (Krassilnikov, N.
A. Actinomycetales. Izvest. Akad. Nauk.
SSSR, Moskau, p. 30, 1941).
Morphology: Sporophores produce long
open spirals. Spores cylindrical or elongated,
1.0 to 1.5 by 0.7 uw, some rounded at ends
and swollen in center.
Agar media: Growth yellow to lemon-yel-
low or dirty yellow, seldom golden yellow.
Pigment insoluble. Aerial mycelium well de-
veloped, velvety, whitish-yellow to brown-
ish-vellow.
Gelatin: Liquefaction medium.
Milk: Coagulation and slow peptonization.
Starch: Hydrolysis weak.
Cellulose: No growth.
Nitrate reduction: Positive.
Sucrose: No inversion.
Antagonistic properties: Weakly antago-
nistic.
femarks: Some strains, such as S. flavo-
viridis, sometimes have a greenish or green-
ish-yellow color instead of a yellow color.
This organism and related forms belong to
the same group as S. griseoflavus and S.
microflavus. The form described by Gause
ct al. (1957) as A. aurini also belongs to this
group.
139. Streptomyces lucensis Arcamone et al.,
1957* (Arcamone, F., Bertazzoli, C., Cane-
A., Ghione, M., and
Microbiol. 4: 119-128,
vazzil, G., DiMarco,
Grein, A.
1957).
Morphology :
Giorn.
Aerial hyphae — long,
branched, and hooked at the tip. Spirals
produced abundantly.
Glycerol-glycine agar: Growth abun-
dant, lemon-yellow. Aerial mycelium gray-
brown. Some soluble pigment produced.
Glucose-asparagine agar: Growth abun-
dant. Aerial mycelium hazel-brown; scanty
clusters of white, short, sterile hyphae. No
soluble pigment.
Potato-glucose agar: Growth abundant,
smooth, yellowish. Aerial mycelium abun-
dant, buff-gray to hazel-brown. Soluble pig-
ment ash-gray, later turning gray-brown.
Yeast-glucose agar: Growth dark brown.
Aerial mycelium whitish. Soluble pigment
dark brown.
Starch agar: Growth abundant, colorless
to yellowish-brown. Aerial mycelium pow-
dery, buff-gray to light brown. No soluble
pigment. Moderate starch hydrolysis.
Oatmeal agar: Growth yellowish and
smooth. Aerial mycelium hazel-brown. No
soluble pigment.
Potato plug: Growth abundant, wrinkled.
Aerial mycelium light gray to hazel-brown.
Plug
culture.
Gelatin: Growth abundant, brown. Aerial
turning
surface turns dark brown around
mycelium white, erayish-brown.
Substrate
)
is strongly darkened within 3
days. No liquefaction.
properties:
antifungal antibiotic, etruscomycin, of the
Antagonistic Produces — an
tetraene type.
Type culture: IMRU 3783.
140. Streptomyces luridus (krassilnikov
et al., 1957) Waksman (Krassilnikov, N. A.,
Koreniako, A. I., Meksina, M. M., Vale-
* Supplemented by personal communication.
238 THE ACTINOMYCETES, Vol. II
dinskaia, L. K., and Vesselov, N. M. Mikro-
biologiya 26: 558-564, 1957).
Morphology: Substrate mycelium mono-
podially branched, 0.7 to 0.8 uw in diameter.
Sporophores produce spirals with 1 to 3
turns. Spores spherical, oval, seldom elon-
gated. Sporulation is generally weak, oc-
curring only on certain media; spiral forma-
tion occurs seldom, largely on synthetic
media with a limited amount of sucrose, and
on starch media.
nitrate agar:
orange. Aerial mycelium white with rose
Sucrose Growth yellow-
tinge.
Nutrient agar: Substrate growth colorless,
free of aerial mycelium. No soluble pigment.
In old cultures, clumps of aerial hyphae may
be formed.
Potato agar: Growth yellow-orange. Aerial
mycelium white with rose tinge. Crystals of
salts deposited along the mycelium in the
substrate.
Milk: Coagulation weak; rapid peptoniza-
tion.
Gelatin: Not liquefied in 10 days.
Starch: Moderate hydrolysis.
Nitrate: Reduced.
Sucrose: Not inverted.
Cellulose: No growth.
Carbon utilization with acid formation:
arabinose, inositol, sorbitol; no acid from
elucose, lactose, rhamnose, xylose, inulin,
inositol, mannitol, or dulcitol.
Antagonistic properties: Produces anti-
bacterial and antiviral (uridin) substances.
Yemarks: Cannot be distinguished from
S. fradiae in its morphological and cultural
properties, but is different in its biochemical
and its antibiotic activities.
141. Streptomyces luteoverticillatus Shin-
obu, 1956 (Shinobu, R. Mem. Osaka Univ.
B (N.S.) 5: 84-93, 1956).
Primary secondary
Morphology: and
verticils produced on various synthetic
media. Spores coccoid to elliptical, about 0.8
u long.
Sucrose nitrate agar: Growth pale brown.
Aerial mycelium cottony, brownish-white
to brown.
Glucose-asparagine agar: Growth thin,
brown. Aerial mycelium cottony, yellow-
white. Soluble pigment pale brown.
Nutrient agar: Growth excellent, deep
brown. Aerial mycelium white to yellow to
green. Soluble pigment deep brown.
Potato plug: Growth heavy, brown. Aerial
mycelium yellow to greenish. Soluble pig-
ment brown.
Milk: Aerial mycelium heavy, brown.
Coagulation uncertain; peptonization strong.
Soluble pigment brown.
Gelatin: Strong liquefaction.
Diastase reaction: Strong.
Tyrosinase reaction: Strong.
Carbon utilization: Fructose, mannitol,
and inositol utilized. Xylose, rhamnose,
sucrose, lactose, and raffinose not utilized.
Habitat: Soil.
Remarks: Resembles S. verticillatus.
142. Streptomyces lydicus DeBoer et al.,
1955 (DeBoer, C., Dietz, A., Silver, W. S.,
and Savage, G. M. Antibiotics Ann. 1955-
1956, p. 886-892).
Morphology: Sporophores long, slightly
coiled at tip. Spores spherical to oval.
Sucrose nitrate agar: Substrate growth
buff-colored. Aerial mycelium white.
Nutrient agar: Some substrate growth.
No aerial mycelium. Soluble pigment yellow-
ish.
Casein-yeast extract-beef agar: Growth
olive-tan. Aerial mycelium gray-white with
flecks of black. Soluble pigment olive-tan.
Glucose-peptone agar: Aerial mycelium
eray-white. Soluble pigment vellow.
Starch agar: Growth good. Aerial myce-
lium pink-gray-white. Hydrolysis good to
excellent.
Gelatin: Some growth. No aerial myce-
lium. Liquefaction positive. Soluble pigment
olive-colored.
DESCRIPTION OF SPECIES OF STREPTOMYCES 239
Milk: Ring around surface. Peptonization
positive.
Nitrate reduction: Positive.
Carbon utilization: Most and
organic acids utilized, but not rhamnose,
inulin, dulecitol, mositol, or the sodium salts
of formic, oxalic, tartaric, and salicylic acids.
Production of H.S: Negative.
Antagonistic properties: Produces an anti-
sugars
biotic, streptolydigin, active against various
bacteria.
Remarks: Related to S. diastaticus.
143. Streptomyces macrosporeus KEttlinger
et al., 1958 (Ettlinger, L., Corbaz, R., and
Hiitter, R. Arch. Mikrobiol. 31: 346, 1958).
Morphology: Sporophores monopodially
branched along the whole axis with open,
irregular spirals. Spores large, 1.7 to 2 by
1.5 to 2 uw; short spines (Plate II m).
Glycerol nitrate agar: Substrate growth
yellow. Aerial mycelium white-yellow to ash-
gray. Soluble pigment golden yellow.
Glucose-asparagine agar: Growth milk-
white. No aerial mycelium. Soluble pigment
whitish-yellow.
Calcium malate agar: Growth yellow.
Aerial mycelium white-yellow to ash-gray.
Soluble pigment white-yellow.
Starch agar: Growth light yellow. Aerial
mycelium white-gray. Good hydrolysis of
starch.
Potato: Growth abundant, light yellow to
golden yellow. Aerial mycelium ash-gray.
Gelatin: Growth sparse. Liquefaction
slow. No soluble pigment.
Milk: Pellicle light yellow
brown. Aerial mycelium white to white-gray.
Coagulation strong; no peptonization.
to yellow-
Antagonistic properties: Produces an
antibiotic, carbomycin.
Habitat: Soil in Madras, India.
144. Streptomyces maculatus (Millard and
Burr, 1926) Waksman and Henrici, 1948
(Millard, W. A. and Burr, 8. Ann. Appl. Biol.
13: 580, 1926).
Morphology: Growth tough, shiny, carti-
laginous. Aerial mycelium rarely produced,
though in certain strains it may frequently
occur, especially when grown on organic
media. Sporophores straight, short. Spores
spherical, 0.5 to 0.6 wu.
Sucrose nitrate agar:
yellow to orange-red; as the culture ages it
Growth orange-
may change to dark green or black. No
aerial mycelium.
Nutrient potato agar: Growth vinaceous-
tawny. Soluble pigment vinaceous-tawny.
Potato: Growth restricted, raised, pinkish.
Aerial mycelium scant, white. Soluble pig-
ment gray to brown.
Gelatin: Growth
slow.
Milk: Growth shght. No coagulation; no
peptonization.
Starch: Hydrolyzed.
Nitrate reduction: Negative.
limited. Liquefaction
Tyrosinase reaction: Negative.
Oxygen requirement: Said to grow well
under anaerobic conditions.
Paraffin: Not utilized.
Temperature: Grows well at 37.5°C.
Habitat: Potato scab and soil.
Type culture: IMRU 3376.
145. Streptomyces madurae — (Vincent,
1894) nov. comb. (Vincent, H. Ann. inst.
Pasteur 8: 129, 1894).
Synonyms: N. babiensis, N. brumpti, N.
madurae, and N. salmonicolor (A. salmoni-
color Millard and Burr, 1926). Baldacci (1944)
listed 17 synonyms.
Strains of this organism were reported by
recently by
Gonzalez Ochoa and Sandoval (1951), to
various investigators, most
form, under certain conditions of culture
and on certain media, such as grain, an
aerial mycelium, with straight or spiral-
shaped sporophores; the spores were cylindri-
cal or oval. This led them to consider this
organism as a Streptomyces. Mariat (1957)
was also of the same opinion. Mackinnon
and Artagaveytia-Allende (1956) consider
240
the generic position of this species as far
from settled.
Morphology: Growth red to red-brown or
pink. In tissues it forms granules consisting
of radiating actinomycosis. Initial branched
mycelium is said to be nonsegmented. Not
acid-fast. Aerial mycelium white and pink
in color.
Glucose-asparagine agar: Growth cream-
colored. Some cultures give reddish pig-
mentation.
Protein media: Growth good, pinkish.
Soluble pigment brown.
Gelatin: Growth glistening, at first white,
then buff to rose or crimson. Soluble pigment
irregular and unpredictable, occasionally red.
Gelatin slowly liquefied.
Milk: No change, or shg¢ht; coagulation
slow, if any; peptonization slow.
Carbon utilization: Utilizes starch, glu-
cose, mannitol, and xylose, but not lactose
or paraffin.
Nitrate reduction: Positive.
Pathogenicity: This property was vari-
ously reported. Topley and Wilson (1946)
stated that this organism causes a local
tissue disease when inoculated under the skin
in guinea pigs. Often reported as not patho-
genic for the usual laboratory animals;
pathogenic for monkeys.
Source: Wide geographical distribution.
Madura foot and other substrates.
146. Streptomyces marginatus (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8S. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight. Spores
oval to spherical, 0.9 by 0.8 uw.
Sucrose nitrate agar: Growth thin, echi-
nate. Aerial mycelium. olive-buff. Soluble
pigment cream-colored.
Nutrient
gray. Aerial mycelium white to whitish-
potato Growth heavy,
agar:
yellow. Soluble pigment light golden brown
to deep golden brown.
Potato: Growth good, raised. Aerial myce-
THE ACTINOMYCETES, Vol. II
lium abundant, buff to olive-buff. Plug at
first gray, later becoming black.
Gelatin: Growth thin, pale olive-gray.
Aerial mycelium abundant, pale gray to
olive-buff. Soluble pigment buff. Liquefac-
tion rapid.
Milk: Growth Aerial mycelium
white. Coagulation and peptonization.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Temperature: Grows well at 387.5°C
Habitat: Potato scab.
good.
147. Streptomyces marinolimosus (Zobell
and Upham, 1944) Waksman (ZoBell, C. E.
and Upham, H. C. Bull. Scripps Inst.
Oceanogr. Univ. Calif. 5: 239-292, 1944).
Morphology: Aerial mycelium consists of
branching filaments with chains of spores.
Spores 0.9 uw in diameter.
Agar media: Growth dull. Aerial myce-
lium white to pinkish, powdery, rough, in
concentric circles. Odor of freshly turned
soil.
Potato: Growth yellow, becoming white,
powdery, raised, rugose. Potato darkened.
Gelatin: Growth in form of flat, yellowish,
circular colonies, with tendency to grow
together. Liquefaction crateriform. Melanin-
negative.
Sea water broth: Growth in form of light
yellow clumps; pellicle produced on surface.
Earthy odor.
Milk: Growth in form of pellicle. Com-
plete peptonization in 20 days.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Production of H.S: Positive.
Source: Marine mud.
Remarks: All differential media were pre-
pared with sea water.
148. Streptomyces marinus (Humm_ and
Shepard, 1946) nov. comb. (Humm, H. J.
and Shepard, IK. S$. Duke Univ. Marine Sta.
Bull. 3: 77, 1946).
Morphology: Sporophores sometimes form
DESCRIPTION OF SPECIES OF STREPTOMYCES
loose spirals. Spores spherical to oval, 0.8
to 1.2 u, produced as a dark gray area in
center of colonies.
Agar media: Growth sparingly branched,
dense, entangled, frequently forming con-
centric rings In response to alternate periods
of light and Aerial mycelium
white. Spores gray to dark gray. No soluble
pigment.
Gelatin: Growth arborescent. Liquefaction
stratiform, slow. Melanin-negative.
Milk: Peptonization
within 1 month.
Starch: Vigorous hydrolysis.
Cellulose: Not attacked.
Chitin and alginic acid: Attacked.
Agar: Slowly digested, softened, not lique-
fied. Growth on agar in culture dish sur-
darkness.
complete, usually
rounded by rather wide, gently sloping
depression. Gelase field relatively wide, with
distinct margin. Irish moss and Hypnea gels
also slowly digested.
Nitrate reduction: Usually negative. In
some media, slight nitrite is produced after
10 days’ incubation, especially if glucose is
present.
Production of H.S: Positive.
Indole: Not formed.
Carbon utilization: Acid produced from
galactose, glucose, fructose, mannose, cel-
lobiose, lactose, maltose, sucrose, and
glycerol. Arabinose, xylose, rhamnose, and
sorbitol utilized without acid production.
No growth with raffinose, salicin, inulin,
dulcitol, inositol, ethyl alcohol, or ethylene
glycol. Utilizes acetic, citric, lactic, propi-
onic, succinic, and 7so-valeric acids. Does not
utilize butyric, gluconic, maleic, malonic,
and oxalic acids.
Habitat: Marine sediments.
149. Streptomyces mashuensis Sawazaki
et al., 1955 (Sawazaki, T., Susuki, 8., Naka-
mura, G., Kawasaki, M., Yamashita, S.,
Isono, K., Anzai, K., Serizawa, Y., and
Sekiyama, Y. J. Antibiotics (Japan) 8A:
44-47, 1955).
241
Morphology: Sporophores straight, no
spirals; numerous primary and secondary
verticils.
Sucrose Growth
nitrate yellow;
reverse yellow-green. Aerial mycelium abun-
agar:
dant, powdery.
Glucose-asparagine agar: Growth pow-
dery, grayish-white, reverse yellow-brown.
No aerial mycelium.
Nutrient
brown.
agar: Growth cream-colored;
reverse No. aerial
soluble pigment.
mycelium. No
Starch agar: Growth marguerite-colored ;
margin cottony, primrose-pink; reverse yel-
low-brown, margin white. Aerial mycelium
white. Strongly diastatic.
Potato: Growth spreading, dark cream-
colored. Aerial mycelium limited. Limited
discoloration of plug.
Gelatin: Growth white. No aerial myce-
lum. Soluble pigment pinkish. Liquefaction
medium.
Nitrate reduction: Negative.
Carbon utilization: Nylose, glucose, su-
crose, trehalose utilized. Rhamnose, raffi-
nose, salicin, mannitol, lactose, arabinose
not utilized.
Antagonistic properties: Produces two
antibiotics, streptomycin and a labile sub-
stance active against mycobacteria, fungi,
and B. subtilis.
Remarks: Okami et al. (1959b) made a
detailed study of this organism. They re-
ported, imstead of the yellow growth on
synthetic media, poor colorless growth.
150. Streptomyces matensis Margalith et
al., 1959 (Margalith, P., Beretta, G., and
Timbal, M. T. Antibiotics & Chemotherapy
9: 71-75, 1959).
Morphology: Sporophores produce verti-
cils, the branches forming spirals. Spores
spherical.
Sucrose nitrate agar: Growth colorless, the
reverse being hyaline to light violet-gray.
Aerial mycelium powdery, gray. Faint
bluish pigment.
242
Glucose-asparagine agar: Growth hyaline
with pinkish reverse. Aerial mycelium pres-
ent. No soluble pigment.
Calcium malate agar:
aerial mycelium.
Nutrient agar: Growth abundant, color-
less. Aerial mycelium whitish. Soluble pig-
Growth poor. No
ment amber.
Starch: Strong hydrolysis.
Potato: Growth abundant. Aerial myce-
lium light gray. No soluble pigment.
Gelatin: Partial liquefaction. No soluble
pigment.
Milk: No coagulation; some peptoniza-
tion.
Nitrate reduction: Negative.
Cellulose: Good growth.
Carbon utilization: Utilizes glucose, su-
crose, lactose, galactose, rhamnose, xylose,
inositol, sodium succinate, and others. Does
not utilize sucrose, raffinose, glycine, or
sodium citrate.
Antagonistic properties: Produces an anti-
bacterial agent, matamycin.
Habitat: Soil.
Remarks: Related to S. noboritoensis and
S. spiralis.
Characteristic properties: Culture said to
be melanin-positive; it produces gray to
violet spore masses.
151. Streptomyces mediocidicus Okami et
al., 1954 (Okami, Y., Utahara, R., Naka-
mura, 8., and Umezawa, H. J. Antibiotics
(Japan) 7A: 98-1038, 1954).
Morphology: Aerial mycelium sometimes
produces verticils, depending on composition
of medium; no spirals.
Glycerol nitrate agar: Growth colorless to
yellowish. Aerial mycelium absent, or white
patches. Soluble pigment absent, or slightly
yellowish brown.
Glucose-asparagine agar: Same as above.
Starch agar: Same as above. Diastatic
action weak or medium.
Nutrient agar: Growth colorless or shghtly
THE ACTINOMYCETES, Vol. II
yellowish. No aerial mycelium. Soluble pig-
ment brownish. Melanin-positive.
Potato: Growth yellowish or light yellow-
ish-brown. Aerial mycelium absent or white.
No soluble pigment.
Gelatin: Growth yellowish-brown. No
aerial mycelium. Soluble pigment brown.
Strong liquefaction.
Blood agar: Growth yellowish-brown to
reddish-brown. Aerial mycelium absent or
white. Hemolytic action strong.
Milk: Surface ring colorless to yellowish.
No aerial mycelium. Soluble pigment
shghtly brownish. Slow coagulation and
peptonization.
Nitrate: No reduction.
Antagonistic properties: Produces an anti-
fungal substance, mediocidin, a polyene of
the hexaene type.
Type culture: IMRU 3777.
152. Streptomyces melanocyclus (Merker,
1911, emend. IWrainsky) Waksman and
Henrici, 1948 (Merker, E. Centr. Bakteriol.
Parasitenk. Abt. II, 31: 589, 1912; Krainsky,
A. ibid. 41: 649-688, 1914).
Morphology: Spores spherical, 0.9 uw.
Agar media: Growth much folded, red.
Aerial mycelium dark brown. Soluble pig-
ment dark brown, turns whole culture red-
brown to almost black with a shade of red.
Calcium malate agar: Colonies small, flat,
orange-red. Aerial mycelium black, occur-
ring along the edges.
Gelatin: Growth poor. Liquefaction rapid.
Milk: Coagulation and rapid peptoniza-
tion.
Starch: Hydrolysis.
Cellulose: Good decomposition; black cir-
cles produced on paper.
Nitrate reduction: Positive.
Sucrose: Inverted.
Pigment: Insoluble in water and in or-
ganic solvents. Considered by Kriss to be
related to the melanins.
Strong effect
Antagonistic properties:
DESCRIPTION OF SPECIES OF STREPTOMYCES
upon various bacteria; some strains show
no activity.
Habitat: Soil.
Remarks: A.
1914) and A.
1928) are related to above species.
melanosporeus (IXrainsky,
melanogenes (Rubentschik
1538. Streptomyces melanogenes Sugawara
and Onuma, 1957 (Sugawara, R. and
Onuma, M. J. Antibiotics (Japan) LOA: 138—
Hee 1957)
Morphology: Sporophores monopodially
branched; no spirals, sometimes slight cur-
vature. Spores cylindrical, 1.7 to 0.8 by
0.9 to 0.5 up.
Sucrose nitrate agar: Growth moist, fold-
ing, colorless to grayish-red-brown; reverse
yellow-orange. Aerial mycelium thin, brown-
ish-white. Soluble pigment brownish-yellow.
Glucose-asparagine agar: Growth colorless
to cream-colored with dark reddish center;
reverse dark yellow-orange. Aerial my-
celium pale grayish-white. Soluble pigment
yellowish-brown.
Calcium malate agar: Growth colorless
to brownish-yellow to grayish-blue-black.
Aerial mycelium yellow-white. Soluble pig-
ment greenish-yellow to brown.
Nutrient agar: Growth cream-colored to
brown. No aerial mycelium. Soluble pigment
reddish-brown.
Potato: Growth folded, colorless to yel-
Aerial mycelium
Soluble pigment
brownish
dark
lowish-brown.
or grayish-white.
yellowish-brown.
Gelatin: Growth colorless to dark brown.
Aerial mycelium white to gray. Soluble pig-
ment pale yellowish-brown. Liquefaction
weak.
Milk: Cream-colored to dark brown ring.
Soluble pigment pinkish-brown.
Blood agar: Growth glistening, vellowish-
gray to dark olive-gray. No aerial mycelium.
Soluble pigment dark brown. Hemolysis
positive.
Antagonistic properties: Produces a mela-
nin-like tumor-inhibiting substance.
243
Remarks: Resembles S. phaeochromo-
genes, S. griseocarneus, and S. cinnamonen-
SUS.
154. Streptomyces michiganensis Corbaz
et al., 1957 (Corbaz, R., Ettlinger, L.,
Keller-Schierlein, W., and Zahner, H. Arch.
Mikrobiol. 26: 192-208, 1957).
Morphology: Sporophores straight, ar-
ranged in sympodially branched clusters;
no spirals. Spores smooth (Pl. II i).
Glycerol nitrate agar: Growth whitish-
yellow. Aerial mycelium velvety, white to
yellowish to greenish-gray.
Calcium malate agar: Growth thin, golden
vellow. Aerial mycelium chalk-white, be-
coming light yellow.
Glucose-asparagine agar: Growth thin,
white to yellow, changing to light yellow-
red. Aerial mycelium velvety, white-yellow.
Glucose-peptone agar: Growth wrinkled,
at first hght brown, then copper-red, finally
reddish-brown. Aerial mycelium velvety,
ereenish-gray. Soluble pigment reddish-
brown.
Gelatin: Pellicle ight brown. Aerial my-
celium powdery, chalk-white. Liquefaction
slow. Soluble pigment brown.
Starch: No hydrolysis.
Potato: Growth light yellow. Aerial my-
celium velvety, white-gray to white-yellow.
Soluble pigment gray-black.
Milk: Pellicle light brown. Aerial myce-
lium sparse. Coagulation and peptonization.
Tyrosinase reaction: Positive.
Antagonistic properties: Produces actino-
mycin X.
Carbon utilization: Xylose,
fructose, galactose, maltose, mannitol, salicin
arabinose,
utilized. Rhamnose, sucrose, lactose, raffi-
nose, inulin not utilized.
Habitat: Soil.
155. Streptomyces microflavus (KXrainsky,
1914) Waksman and Henrici, 1948 (Krain-
sky, A. Centr. Bakteriol. Parasitenk. Abt.
II., 41: 686, 1914).
244 THE ACTINOMYCETES, Vol. II
Morphology: Spores spherical to rod-
shaped, often produced in pairs or in chains,
2-Ov byt Zo aa:
Calcium malate agar: Colonies minute,
yellow. No aerial mycelium.
Glucose-asparagine Aerial
lium produced late (12 days), rose-yellow.
Nutrient agar: Colonies yellow. Aerial
mycelium produced late, yellowish-rose.
Potato: Growth yellowish, slimy mass.
No aerial mycelium. Melanin-negative.
Gelatin: Colonies small, yellowish. Lique-
agar: myce-
faction rapid.
Milk: Rapid coagulation and peptoniza-
tion.
Invertase: Negative.
Starch: Diastatic action strong.
Cellulose: Growth scant, white.
Nitrate reduction: Positive.
Production of HS: Negative.
Antagonistic properties: Said to produce
a form of streptothricin.
Habitat: Soil.
Remarks: According to Ettlinger et al.
(1958), this organism belongs to the S.
griseus series.
Type culture: IMRU 3332; ATCC 13,281.
156. Streptomyces mirabilis Ruschmann,
1952 (Ruschmann, G. Pharmazie 7: 542-
550, 639-648, 823-831, 1952).
Morphology: Sporophores straight, with-
out spirals or curvature.
Agar media: Aerial mycelium white,
cottony.
Nutrient agar: Growth
slimy surface. No aerial mycelium.
Glucose agar: Growth grayish-brown. No
poor, forming
aerial mycelium. Soluble pigment brown.
Potato: Growth good, lichenoid. Soluble
pigment dark brown to black.
Gelatin: Good flaky growth. Rapid lique-
faction. Soluble pigment dark brown to
black.
Milk: Surface growth covered with white,
fluffy Coagulation and
aerial mycelium.
peptonization positive. Liquefied
colored black.
Fats: Ready utilization.
Temperature: Optimum 29°C. No growth
portion
(> ay AO}
at o¢
Antagonistic properties: Antagonistic ef-
fect strongest in freshly isolated cultures.
Property lost on cultivation; activity lost
first against gram-negative, rod-shaped bac-
teria, coccl remaining most sensitive. Pro-
duces antibiotic miramycin.
temarks: Highly proteolytic and lipoly-
tic. Grows best on complex organic media,
at slightly acid reaction, pH 6.0 to 6.6.
157. Streptomyces mitakaensis Arai et al.,
1958 (Arai, M., Karasawa, K., Nakamura,
S., Yonehara, H., and Umezawa, H. J.
Antibiotics (Japan) 11A: 14-20, 1958).
Morphology : Sporophores short,
branched; spirals produced. Spores spheri-
eal, 1.2: to 1.5: a:
Sucrose nitrate agar: Growth good, color-
less or white to dark yellowish-brown. Aerial
mycelium powdery, abundant, light gull-
eray. No soluble pigment.
Glucose-asparagine agar: Growth good,
white to brown; becomes dark
brown. Aerial mycelium powdery, abundant,
gray. No soluble pigment.
Glycerol citrate Growth
colorless or white to brownish-white; later
brownish-yellow. Aerial mycelium powdery,
abundant, whitish-gray in center, gray in
the edges. No soluble pigment.
Nutrient agar: Growth good, colorless to
reverse
agar: good,
pale yellowish-brown. Aerial mycelium pow-
dery, abundant, white, with or without
eray parts. No soluble pigment.
Starch agar: Growth good, colorless to
pale yellowish-brown. Aerial mycelium pow-
dery, abundant, light gray in center, gray
at the edges. No soluble pigment. Hydroly-
sis strong.
Potato plug: Growth
eream-buff. Aerial mycelium poorly devel-
good, wrinkled,
DESCRIPTION OF SPECIES OF STREPTOMYCES
oped, white or grayish-white. No soluble
pigment.
Gelatin: Growth colorless. Aerial myce-
lium pale gull-gray. Soluble pigment absent,
yellow. Liquefaction moderate.
Milk: Growth good, colorless to white.
Peptonization and coagulation rapid.
Nitrate reduction: Negative.
Antagonistic properties: Produces — an
antibiotic, mikamycin, active against gram-
positive and acid-fast bacteria.
158. Streptomyces = murinus Frommer
(Frommer, W. Arch. Mikrobiol. 32: 198,
1959).
Morphology: Sporophores small, mono-
podially branched, tree-like; spirals com-
pact, with 1 to 3 turns.
Glycerol nitrate agar: Growth greenish-
yellow. Aerial mycelium thin, white. Soluble
pigment greenish-yellow.
Glycerol-glycine yellow
to brown-yellow. Aerial mycelium white-
gray to gray-brown. Soluble pigment golden
agar: Growth
yellow.
0 Growth yellow,
occasionally brown-violet. Aerial mycelium
powdery white to gray-white. Soluble pig-
Glucose-asparagine agar:
ment yellow to greenish-yellow.
Calcium malate agar: Growth colorless.
Aerial mycelium white. No soluble pigment.
Nutrient agar: Growth yellow to green-
ish-yellow. Aerial mycelium lacking or
white. Soluble pigment lacking or golden
yellow.
Starch media: Growth colorless to yellow-
ish. Aerial mycelium gray-brown. No hy-
drolysis after 10 days.
Potato: Growth abundant, golden brown.
Aerial mycelium cream-colored to yellow.
Soluble pigment questionable.
Gelatin: Growth abundant, golden yellow.
Aerial mycelium eray.
Soluble pigment yellow to golden yellow.
Liquefaction limited. Melanin-negative.
Milk: Growth abundant, golden yellow to
cream-colored to
245
Aerial mycelium powdery,
cream-colored. Questionable coagulation and
vellow-brown.
liquefaction.
Cellulose: Growth weak. Aerial mycelium
gray-brown. Soluble pigment yellowish.
Antagonistic properties: Produces actino-
mycin.
159. Streptomyces naganisht Yamaguchi
and Saburi, 1955 (Yamaguchi, T. and
Saburi, Y. J. Gen. Appl. Microbiol. 1: 201-
230 L955):
Morphology: Sporophores straight with
many compact spirals and a few open
spirals; spores oval to short rods, 0.8 to 1.4
by 0.5 to 0.7 up.
Sucrose nitrate agar: Growth colorless,
thin. Aerial mycelium powdery, at first
white, later colored buff. No soluble pig-
ment.
Calcium malate agar: Growth is at first
pinkish-white to pinkish-gray, later becom-
ing whitish-brown. Aerial mycelium whitish.
Soluble pigment light pink, but soon disap-
pears.
Nutrient agar: Growth at first colorless
to dark cream, later becoming yellowish-
brown to brown. No aerial mycelium. Solu-
ble pigment light brown. Melanin-negative.
Starch agar: Growth colorless to creamy
with reddish-purple portion. Aerial myce-
lium abundant, white or smoke-gray to light
drab. Soluble pigment absent or faint pink.
Good hydrolysis.
Potato: Growth vigorous, at first yellow-
ish-gray. Aerial mycelium white to grayish-
white. Soluble pigment deep purple to black.
Gelatin: Growth dark brown with some
tint of olive. Soluble pigment deep brown
and a more diffusible yellowish-green. Lique-
faction moderate.
Milk: Growth vigorous, yellowish-brown,
with white aerial mycelium along the glass.
Soluble pigment hght
reddish-brown. Coagulation and peptoniza-
brown, sometimes
tion.
246
Carbon utilization: Utilizes p-xylose,
L-arabinose, L-rhamnose, D-galactose, lactose,
raffinose, mannitol, inositol, salicin, acetate,
citrate, and succinate; does not utilize su-
crose, inulin, sorbitol, or cellulose.
Antagonistic properties: Active against
gram-positive and acid-fast bacteria, fung),
and trichomonads.
temarks: Related to S. antimycoticus.
160. Streptomyces narbonensis Corbaz_ et
al., 1955 (Corbaz, R., Ettlinger, L., Gau-
mann, E., Keller, W., Kradolfer, F., Ky-
burz, E., Neipp, L., Prelog, V., Reusser, R.,
and Zahner, H. Helv. Chim. Acta 38: 935-
942, 1955).
Morphology: Sporophores
spirals. Spores smooth, cylindrical, 0.8 to
Lil byaO2760.0.9 ju:
Glycerol nitrate agar: Growth thin, color-
less to yellowish-brown. Aerial mycelium
velvety, whitish-gray. No soluble pigment.
Glucose-asparagine agar: Growth thin, at
first colorless, then yellowish-brown. Aerial
mycelium sparse, chalk-white. No soluble
straight; no
pigment.
Calcium malate agar: Growth colorless.
as
FiGguRE 43. isolate AA 877, re-
sembling S.nelropsis, showing character of verticils
Hyphae of
of sporogenous branches (Reproduced from: Dug-
gar, B. M. etal. Ann. N. Y. Acad. Sei. 60: 85, 1954).
THE ACTINOMYCETES, Vol. II
Aerial mycelium white-gray. No soluble
pigment.
Glucose-peptone agar: Growth light yel-
lowish, punctiform. Aerial mycelium pro-
duced late, powdery, gray-white. No soluble
pigment.
Nutrient agar: Growth punctiform, yel-
lowish. No aerial mycelium. No soluble
pigment. Melanin-negative.
Starch agar: Growth thin, colorless to
vellowish. Aerial mycelium powdery, white.
No soluble pigment. Hydrolysis good.
Gelatin: Growth yellowish-white. Aerial
mycelium snow-white. Soluble pigment light
reddish-brown. Liquefaction slow.
Potato: Growth lichenoid, bluish-gray to
reddish-gray. No aerial mycelium. Soluble
pigment dark brown.
Milk: Surface ring whitish-yellow. Pep-
tonization without coagulation.
Production of H.S: Positive.
Antagonistic properties: Produces basic
antibiotic, narbomycin, related to picromy-
cin.
Carbon utilization: Utilizes xylose, arabi-
nose, rhamnose, fructose, galactose, sac-
charose, maltose, raffinose, inulin, salicin,
sodium acetate. Does not utilize mannitol,
sorbitol, dulcitol, mesoinositol.
Habitat: Soil.
femarks: Ettlinger et al. (1958) consider
this organism as belonging to S. olzvaceus.
161. Streptomyces netropsis Finlay and
Sobin, 1952 (Finlay, A. C. and Sobin, B. A.
U.S. 2.586.762. 1952).
Morphology: Sporophores in form of ver-
ticils or terminal clusters on tips of short
hyphae (Fig. 43). Spores short, cylindrical,
0.7 by 1.8 uw, smooth (Pl. I a).
Sucrose nitrate agar: Growth thin, pale
olive-buff. Aerial mycelium pale vinaceous-
fawn. No soluble pigment.
Glucose-asparagine agar:
ate, wrinkled. Aerial mycelium white. Solu-
Growth moder-
ble pigment brown.
Calcium malate agar: Growth moderate,
DESCRIPTION OF SPECIES OF STREPTOMYCES 247
cream to buff. Aerial mycelium white. No
soluble pigment.
Nutrient agar: Growth moderate to good,
light brown. Aerial mycelium white. Solu-
ble pigment light brown.
Starch agar: Growth moderate, thin; pale
olive-buff reverse. Aerial mycelium white.
No soluble pigment. Strong hydrolysis.
Potato: Growth poor, waxy, wrinkled,
brown. No aerial mycelium. Soluble pig-
ment dark brown.
Gelatin: Moderate surface growth. Aerial
mycelium white. Soluble pigment dark
brown. No liquefaction.
Milk: Growth poor. No peptonization.
Nitrate reduction: Negative.
Production of HS: Variable.
Antagonistic properties: Produces a basic
antibiotic, netropsin.
Remarks: Ettlinger et al. (1958) report
this organism to be melanin-negative. They
also consider S. cznnamomeus as closely re-
lated.
162. Streptomyces niger (Rossi-Doria,
1891; emend. Krassilnikov, 1949) Waksman
(Rossi-Doria, E. Ann. igiene, 1: 399-43
1891; Krassilnikov, N. A. Actinomycetales.
Izvest. Akad. Nauk. SSSR, Moskau, p. 53,
1941).
Morphology: Substrate growth of soft
consistency. Aerial mycelium produced only
on potato and synthetic agar. Sporophores
formed only seldom; open spirals, with 3 to
5 turns. Spores oval.
Synthetic agar: Growth black. Aerial
mycelium dark gray. No soluble pigment.
Nutrient agar: Growth black. Soluble
pigment brown.
Gelatin: Slow liquefaction,
Melanin-negative (?).
Milk: No change.
Starch: No growth.
Cellulose: No growth.
Nitrate reduction: Negative.
Sucrose: No inversion.
Temperature: Optimum 25-30°C.
in 30 days.
Antagonistic properties: None.
Remarks: This is a very unstable species
which dies out rapidly. It easily mutates,
giving rise to colorless cultures, producing
no aerial mycelium. It appears
transition form, if not a true Nocardia. A.
niger aromaticus Berestnew and A. nigrificans
(Kriiger) Wollenweber are listed by Krassil-
nikov as varieties of A. niger. In view of the
formation of a soluble brown substance on
to be a
certain protein media, this organism may
belong to one of the chromogenic groups.
163. Streptomyces nigrifaciens Waksman,
1919 (Waksman, 8. A. Soil Sci. 8: 167-168,
1919).
Morphology: Sporophores branching with
tendency to curl; no true spirals. Spores
oval-shaped to elliptical.
Sucrose nitrate agar: Growth colorless.
Aerial mycelium thin, gray. No soluble pig-
ment.
Glucose-asparagine agar: Growth cream-
colored. Aerial mycelium mouse-gray with
white patches.
Nutrient agar: Growth thin, cream-col-
ored. Aerial mycelium gray. Soluble pigment
brown.
Starch agar:
yellow. Aerial mycelium light
Hydrolysis imperfect.
Ege media: Growth abundant, dark
brown. No aerial mycelium. Purplish zone
around growth.
cream-colored to
buff-gray.
Growth
Potato: Growth gray becoming dark.
Aerial mycelium white, appearing late.
Soluble pigment black.
Gelatin: Growth cream-colored to brown-
ish. Aerial mycelium white. Soluble pigment
brown. Liquefaction slow.
Milk: Surface growth dark brown. Aerial
mycelium white. Coagulation and slow pep-
tonization.
Nitrate reduction: Positive.
Sucrose: No inversion.
Cellulose: No growth.
Habitat: Pineapple soil in Hawaii.
248
Remarks: This organism had been de-
scribed by Waksman (1919) as Actinomy-
ces 145, but never named before.
Type culture: IMRU 3067.
164. Streptomyces nitrificans Schatz et al.,
1954 (Schatz, A., Isenberg, H. D., Angrist,
A. A., and Schatz, V. J. Bacteriol. 68: 1-4,
1954).
Morphology: Sporulating hyphae straight,
branched.
Most solid and liquid media: Growth
gray, with a pink to buff reverse. No solu-
ble pigment.
Blood agar: Growth brick-red. No hemoly-
sis.
Potato: Growth wrinkled.
Nitrate reduction: Positive.
Milk: No coagulation; slow peptonization.
Starch: Hydrolysis.
Gelatin: No liquefaction.
Cellulose: Not attacked.
Remarks: S. nitrificans grows well on a
variety of substrates, such as ethyl carbam-
ate. With ammonia providing nitrogen in the
basal medium, glucose, sucrose, mannitol,
sorbitol, glycerol, ethanol, n-propanol, ace-
tate, lactate, succinate, fumarate, and citrate
permitted good growth. In a glucose con-
taining medium, ammonia, nitrite, nitrate,
urea, and guanidine were satisfactory sources
of nitrogen. Several amino acids, purines,
and miscellaneous other nitrogenous com-
pounds, supped alone or with glucose in
the basal medium, supported growth.
The organism grew as well on carbamate
when first isolated from a carbamate-enrich-
ment culture as it did after serial transfer
over a 2-year period on various simple and
complex media containing no carbamate.
In addition to its apparently unique abil-
itv to grow on carbamate as sole substrate,
this culture also produced nitrite from car-
bamate. It did not oxidize the carbamate
nitrogen beyond the nitrite stage. Hirsch
(1960) considers this organism as a Nocardia
THE ACTINOMYCETES, Vol. II
(N. nitrificans) capable of utilizing petro-
leum.
165. Streptomyces nitrosporeus Okami,
1952 (Okami, Y. J. Antibiotics (Japan) 5:
477-480, 1952).
Morphology: Aerial mycelium. straight,
formed in clusters or tufts. Spores elliptical
to oval.
Sucrose nitrate agar: Substrate growth
colorless, grayish. Aerial mycelium blackish-
oray.
Gelatin: Limited growth in liquefied zone.
Liquefaction rapid. Soluble pigment yellow-
ish-brown.
Milk: Growth cream-colored to brownish.
Strong coagulation and peptonization.
Starch: Strong hydrolysis.
Cellulose: Attacked.
Nitrate reduction: Vigorous.
Tyrosinase reaction: Negative.
Production of HoS: None.
LoefHler’s media: Growth thin.
Aerial mycelium gray. Soluble pigment
limited. Rapid liquefaction of serum.
Carbon utilization: Utilizes arabinose,
galactose, glucose, maltose, rhamnose, xy-
serum
lose, and glycerol; does not utilize sucrose,
fructose, inulin, lactose, mannitol, raffinose,
or sorbitol.
Antagonistic properties: Produces an anti-
biotic, nitrosporin (proactinomycin?).
Habitat: Soil in Japan.
Remarks: Resembles S. griseolus and S.
cellulosae.
Type culture: IMRU 3728; ATCC 12,769.
166. Streptomyces niveoruber Ettlinger et
al., 1958 ‘(Bttlinger, L., Corbaz,.-Resand
Hitter, R. Arch. Mikrobiol. 31: 350, 1958).
Morphology: Long, straight sporophores,
monopodially branched, forming open, regu-
lar spirals. Spores smooth (PI. II k).
Glycerol nitrate agar: Growth hght yellow
or carmine-red. Aerial mycelium sparse,
chalk-white.
Glucose-peptone agar: Growth white-yel-
DESCRIPTION OF SPECIES OF STREPTOMYCES
low to carmine-red. Aerial mycelium abun-
dant, white. Soluble pigment
carmine-red.
Calcium malate agar: Growth light yellow
to ight carmine. No aerial mycelium.
Starch agar: Growth light yellow-red to
carmine-red. Aerial
white. Limited hydrolysis.
Potato: Growth light brown. Aerial myce-
lium powdery, chalk-white. Melanin-nega-
tive.
Gelatin: Growth carmine-red. Aerial my-
celium sparse. Trace of liquefaction. No
soluble pigment.
Milk: Pellicle hight yellow. Aerial myce-
lium sparse. Coagulation limited; no pep-
tonization.
Antagonistic properties:
biotic cimerubin.
Habitat: Soils in England and Germany.
somewhat
mycelium abundant,
Produces anti-
167. Streptomyces niveus Smith et al., 1956
(Smith, C. G., Dietz, A., Sokolski, W. T.,
and Savage, G. M. Antibiotics & Chemo-
therapy 6: 135-142, 1956).
Morphology: Sporophores straight at the
base, corkscrew-coiled at tip, occur in clus-
ters and bear oblong spores.
Sucrose nitrate agar: Growth cream-col-
ored. Aerial mycelium white. Soluble pig-
ment yellow.
Calcium malate agar: Growth cream-
colored. Aerial mycelium white. Soluble
pigment yellow.
Nutrient agar: Growth cream-colored.
Aerial mycelium trace, gray-white. Soluble
pigment yellow.
Gelatin: Growth good. Liquefaction par-
tial. No soluble pigment.
Nutrient starch agar: Growth vellow.
Aerial mycelium cream-pink. Hydrolysis
good.
Tyrosine agar: Soluble yellow pigment.
Milk: Ring on surface; floeculent growth
at bottom. Positive peptonization.
Production of H.S: Negative.
Carbon utilization: b-xylose, b-arabinose,
249
rhamnose, D-fructose, D-galactose, D-glucose,
D-mannose, maltose, sucrose, lactose, cello-
biose, raffinose, dextrin, inulin, soluble starch,
glycerol, duleitol, bD-mannitol, b-sorbitol, im-
ositol, salicin, sodium formate, sodium ox-
alate, sodium tartrate, sodium acetate, so-
dium citrate, and sodium succinate utilized.
Phenol, cresol, and sodium salicylate not
utilized.
Nitrate: No reduction.
Antagonistic properties: Produces strepto-
nivicin, a form of novobiocin.
Habitat: Soil.
Remarks: According to Kuroya et al.
(1958), this organism is related if not identi-
cal to S. griseoflavus.
168. Streptomyces noboritoensis Isono. et
al., 1957 (Isono, K., Yamashita, S., Tomi-
yama, Y., Suzuki, S., and Sakai, H. J. Anti-
biotics (Japan) 10A: 21-30, 1957).
Morphology: Aerial mycelium long and
wavy; no regular spirals.
nitrate Growth
Aerial mycelium slight. Soluble pigment
Sucrose agar: colorless.
absent or pale yellow.
Glucose-asparagine
dark
pale violet-gray.
slightly brownish.
Nutrient agar: Growth flat, pale gray,
smooth and restricted. No aerial mycelium.
Soluble pigment dark red-brown.
Starch agar: Growth dry, wrinkled, pale
agar: Growth pale
brown. Aerial mycelium
No soluble pigment or
brown. to
grayish-brown. Aerial mycelium pale-gray,
cottony. Weak diastatic action.
Gelatin: Growth dark brown. Soluble pig-
ment dark brown. Liquefaction absent or
slight.
Potato: Growth flat, wrinkled, black.
Aerial mycelium grayish-white in some
strains. Color of plug black.
Carbon utilization: Glucose, lactose, man-
nitol, trehalose, and raffinose well utilized.
Utilization of arabinose, inositol, salicin,
and xylose limited. Rhamnose and sucrose
not utilized.
250
Antagonistic properties: Produces anti-
biotic homomycin-hygromycin.
Remarks: S. noboritoensis belongs to the
group of chromogenic actinomycetes closely
related to S. cinnamonensis, S. flavochromo-
genes, S. phaeochromogenes, S. aureus, and
S. tanashiensis. They differ in spiral forma-
tion, pigmentation on synthetic media, ni-
trate reduction, and production of anti-
biotics.
169. Streptomyces nodosus Trejo, W. nov.
Sp:
Morphology: Aerial mycelium forms open
and closed spirals, the latter predominating
as tightly knotted coils. Spores spherical to
oval, 0.5 to 1.0 by 1.0 p.
Sucrose nitrate agar: Growth white-green-
ish. Aerial mycelium pearl-gray to dawn-
eray.
Nutrient agar: Substrate growth scant.
No aerial mycelium. No soluble pigment.
Melanin-negative.
Oatmeal agar: Growth black with a buff
margin. Aerial mycelium deep olive-gray.
Reverse: olivaceus to black with a peripheral
ring of cream-buff to chamois. No soluble
pigment.
Potato: Growth buff. Aerial mycelium
light olive-gray. No darkening of plug.
Milk: Rapidly peptonized.
Gelatin: Rapidly hydrolyzed.
Nitrate reduction: Positive.
Starch: Strong hydrolysis.
Tyrosine: Utilized with no melanin forma-
tion.
Carbon utilization: Utilizes mannitol, in-
ositol, rhamnose, xylose, D-fructose, treha-
lose, and melibiose. Does not utilize adoni-
tol, sorbitol, arabinose, cellulose, sucrose,
lactose, sodium acetate, esculin, or dextrin.
Antagonistic properties: Produces an anti-
fungal antibiotic, amphotericin.
* Personal communication from Squibb Insti-
tute for Medical Research (1958).
THE ACTINOMYCETES, Vol. II
Source: Isolated from soil in South
America.
Remarks: This culture was specially de-
scribed for this treatise. It appears to be
closely related to S. rutgersensis.
170. Streptomyces noursei Hazen and
Brown, 1950 (Hazen, E. L. and Brown, R.
Science 112: 423, 1950; Proc. Soc. Exptl.
Biol. Med 76: 93, 1951; Science 117: 609,
1953).
Morphology: Sporophores produced as
side branches of sterile aerial hyphae; occa-
sionally produce open spirals and, according
to Ettlinger et al. (1958), also some verticils.
Spores round to oval, with thin long spines
CPE ey
Sucrose nitrate agar: Growth scanty, col-
orless, flat. No aerial mycelium.
Glucose-asparagine agar: Growth wrin-
kled, tan-colored, with gray and white knob-
like projections. Reverse of growth dark
eray. Aerial mycelium white, then reddish-
eray, finally ash-gray; limited shell-pink
diffusible pigment.
Glucose-peptone agar: Growth good,
folded, brown. Aerial mycelium white, turn-
ing gray. Soluble pigment brown or pome-
eranate-purple.
Starch agar: Growth in form of discrete
colonies. Aerial mycelium white in center,
periphery colorless and embedded. Hydroly-
Sis.
Potato: Growth folded. Aerial mycelium
chalky white. At 35-36°C a reddish-purple
pigment is formed.
Gelatin: Rapid
negative.
Milk: Coagulation, followed by peptoni-
zation.
Cellulose: Growth poor.
Nitrate: Traces of nitrite produced.
Production of HS: Negative.
Blood agar: Growth consists of convex,
lobate colonies, with central perforation.
Aerial mycelium heavy, chalky white. No
hemolysis, but darkening of blood.
liquefaction. Melanin-
DESCRIPTION OF SPECIES OF STREPTOMYCES
Antagonistic properties: Produces an
antifungal agent, nystatin.
Type culture: IMRU 3771.
171. Streptomyces novaecaesareae (Waks-
man and Curtis, 1916) Waksman and Hen-
rici, 1948 (Waksman, 8. A. and Curtis, R. E.
Soil Sci. 1: 111, 1916, 8: 158, 1919).
Morphology: Aerial mycelium forms both
straight and spiral (dextrorse) sporophores.
Spores oval to elongate.
Sucrose nitrate agar: Growth gray, be-
coming bluish, glossy, much wrinkled. Aerial
mycelium white, appears late. Soluble pur-
ple pigment formed.
Glucose-asparagine agar:
stricted, gray, becoming red.
Nutrient agar: Growth thin, cream-col-
ored.
Potato: Growth wrinkled, cream-colored,
turning yellowish. Melanin-negative.
Gelatin: Surface colonies small, cream-
colored. Liquefaction slow.
Milk: Gray ring. Coagulation slow; pep-
tonization slow.
Starch agar: Colonies restricted, circular,
bluish-violet. Positive hydrolysis.
Nitrate reduction: Positive.
Production of H.S: Negative.
Temperature: Optimum, 37°C.
Antagonistic properties: Negative.
Remarks: At first this organism was des-
ignated as A. violaceus-caesari. This species
is considered by Krassilnikov as synonymous
with A. violaceus (Rossi-Doria) Gasperini.
It appears to be related to S. violaceoruber.
Growth — re-
172. Streptomyces odorifer — (Rullman
emend. Lachner-Sandoval, 1898) Waksman
(Lachner-Sandoval, V. Ueber Strahlenpilze.
Strassburg, 1898).
Morphology: Sporophores long, straight,
branching, forming spirals. Spores spherical.
Sucrose nitrate agar: Growth cream-col-
ored, with trace of brown. Aerial mycelium
abundant, cream-colored.
Glucose-asparagine agar: Growth cream-
251
colored to brownish. Aerial mycelium abun-
dant, cream-colored. Soluble pigment faint
brownish.
Nutrient agar: Growth folded, brown.
Aerial mycelium white around edge. Soluble
pigment faint brown.
Starch agar:
brown. Aerial mycelium abundant, cream-
colored to straw-colored. No soluble pig-
ment. Hydrolysis good.
Growth cream-colored to
Maltose-peptone agar: Foulerton and
Price-Jones (1902) described growth as
“raised, drab-colored, semi-translucent, the
surface becoming reticulated; soluble pig-
ment deep brown; gelatin liquefied, with
light brown pigmentation.”
Potato: Growth folded, brownish. Aerial
mycelium cream-colored. Soluble pigment
faint brown.
Gelatin: Surface ring cream-colored. Ae-
rial mycelium thin, white. No soluble pig-
ment. Liquefaction slow.
Milk: Surface ring colorless to brownish.
No aerial mycelium. No coagulation; some
peptonization.
Cellulose: Good growth.
Sucrose: Inversion.
Paraffin and fats: Good growth.
Nitrate reduction: Positive.
Production of HS: Negative.
Odor: Strong, characteristic of soil.
Antagonistic properties: Some strains give
positive effects, others are negative.
Habitat: Soil.
Type culture: IMRU 3334.
173. Streptomyces oidiosporus (Krassilni-
kov, 1941) Waksman (Krassilnikov, N. A.
Actinomycetales. Izvest. Akad. Nauk.
SSSR, Moskau, p. 23, 1941).
Morphology: Sporophores — straight or
wavy, never forming spirals; short or long,
frequently forming broom-shaped structures.
Spores 1.0 to 1.8 by 0.5 to 1.0 u, frequently
appearing as double cocci or segmented
spores (oidiospores).
Agar media: Growth red or rose to pale;
252 THE ACTINOMYCETES, Vol. II
pigment insoluble in medium. Aerial myce-
lium poorly developed, velvety, rose-white.
Gelatin: Aerial mycelium weakly devel-
oped, frequently lacking; aerial hyphae
short, rose-white. Liquefaction weak.
Milk: No coagulation; peptonization posi-
tive.
Starch: Rapid hydrolysis of starch.
Cellulose: No growth.
Nitrate reduction: Positive.
properties:
positive
None. Jolly
effects for his
Antagonistic
(1956) obtained
strain.
Habitat: Rarely found in soil.
Remarks: The resembles SS.
ruber and S. longispororuber. Some strains
were obtained as variants of Nocardia
rubra. Jolly (1956) reported the isolation of
a strain of S. otdiosporus from an Italian
organism
soil.
174. Streptomyces olivaceus (Waksman,
1919) Waksman and Henrici, 1948 (Waks-
man, S. A. Soil Sci. 8: 168, 1919).
This organism was first described as
strain No. 206 by Waksman (1919). It was
used by Jensen (1930) for comparison with
his own isolates. It was studied more re-
cently by Shinobu (1958) and Ettlinger
et al. (1958).
Morphology: § Sporophores — branched
monopodially, straight or somewhat wavy;
no true spirals on most media; a few long,
open spirals on calcium malate agar. Spores
spherical and oval, 0.8 to 1.2 uw; surface
smooth (Pl. III).
Sucrose nitrate agar: Growth abundant,
yellow to olive-ocher, reverse yellow to al-
most black. Aerial mycelium ash-gray to
light drab.
Glucose-asparagine agar: Growth yellow
to light olive to olive-gray. Aerial mycelium
light olive-gray to light brownish-gray with
greenish tinge. No soluble pigment.
Calcium malate agar: Growth greenish-
yellow to yellow. Aerial mycelium yellowish-
white to yellowish-gray. Soluble pigment
yellow.
Nutrient agar: Growth white, glistening.
No soluble pigment.
Starch agar: Growth brownish-yellow to
yellowish-green. Aerial mycelium brownish-
white. Hydrolysis strong.
Potato: Growth abundant, much wrin-
kled, elevated, gray, turning sulfur-yellow
on edge. Melanin-negative.
Gelatin: Liquefaction rapid. No soluble
pigment.
Milk: Growth faint, pinkish; coagulation
and peptonization rapid.
Cellulose: Growth good.
Mannase: Reaction strong, according to
Shinobu (1958).
Nitrate reduction: Positive.
Production of HS: Negative.
Tyrosinase reaction: Although this organ-
ism has been considered as melanin-nega-
tive, Shinobu (1958) reported a positive
reaction.
Temperature: Optimum 25°C.
Carbon According to Shinobu
(1958), S. olivaceus rapidly utilizes xylose,
rhamnose, fructose,
sources:
galactose, sucrose, lac-
tose, and mannitol; slow utilization: treha-
lose, raffinose, and inositol.
Antagonistic properties: Various strains
produce a variety of antibiotics, including
streptomycin, olivacein, and granaticin.
Habitat: Very common in soil.
Remarks: Krassilnikov (1949) placed the
organism in the A. flavus group. Ettlinger
et al. (1958) considered the following organ-
isms as belonging to S. olzvaceus: S. felleus,
S. flavus, S. griseolus, S. halstedii, S. nar-
bonensis, S. scabies (sic), and S. verne.
Type culture: IMRU 3335.
175. Streptomyces olivochromogenes (Waks-
man, 1919) Waksman and Henrici, 1948
(Waksman, S. A. Actinomyces No. 205, Soil
Sci. 8: 106, 1919).
Morphology: Sporophores form numerous
closed spirals. Spores oval or elliptical.
DESCRIPTION OF SPECIES OF STREPTOMYCES
nitrate Growth white,
spreading. Aerial mycelium ash-gray with
brownish tinge. No soluble pigment.
Glycerol malate agar: Growth colorless.
Aerial mycelium light grayish-olive to dark
oray.
Glucose-asparagine agar: Growth abun-
dant, natal-brown to almost black. Aerial
mycelium white with gray tinge. Soluble
Sucrose agar:
pigment brownish.
Nutrient agar: Growth wrinkled, brown,
becoming gray-green. Aerial mycelium
white. Soluble pigment brown.
Starch agar: Growth transparent, spread-
ing. Aerial mycelium buff-gray. Rapid hy-
drolysis.
Potato: Colonies small, wrinkled, black.
No aerial mycelium. Soluble pigment black.
Gelatin: Surface growth cream-colored,
spreading. Aerial mycelium white. Soluble
pigment dark brown to deep olive-green.
Slow liquefaction.
Milk: Dark brown ring. Coagulation and
peptonization.
Cellulose: Growth faint.
Nitrate reduction: Faint reduction to ni-
trite.
Sucrose: Invertase positive with good
growth.
Temperature: Optimum, 37°C.
Antagonistic properties: Positive.
Habitat: Soil, water, river mud.
Remarks: Ettlinger et al. (1958) consid-
ered this organism as a strain of S. griseus.
KrassilInikov (1949) considered it as a vari-
ety of A. chromogenes.
176. Streptomyces olivoreticuli Arai et al.,
1957 (Arai, T., Nakada, T., and Suzuki,
M. Antibiotics & Chemotherapy 7: 435-442,
1957).
Morphology: Sporophores form primary
and secondary verticils; secondary may also
be formed as tip clusters. Spores spherical
to oval.
Sucrose nitrate agar: Growth thin, yellow
to brown. Aerial mycelium scant, later be-
253
coming cottony, white with yellowish tinge.
Soluble pigment faint brown or absent.
Glucose-asparagine agar: Growth thin,
hight brown to olive-drab. Aerial mycelium
cottony, white with faint yellow to grayish-
pink tinge.
Nutrient agar: Growth limited, brownish.
Aerial mycelium grayish-white. Soluble pig-
ment light brown.
Gelatin: Surface growth poor. Liquefac-
tion slow, later becoming rapid. Soluble pig-
ment brown.
Blood agar: Strong hemolysis.
Potato: Growth wrinkled, dark brown.
Aerial mycelium abundant, powdery, cream-
colored to tea-green. Soluble pigment brown.
Milk: Ring on surface brown. Coagulation
with limited peptonization. Soluble pigment
brown.
Nitrate reduction: Negative.
Starch: Hydrolysis.
Cellulose: No decomposition.
Antagonistic properties: Produces anti-
biotic viomycin.
177. Streptomyces olivoverticillatus —Shi-
nobu, 1956 (Shinobu, R. Mem. Osaka Univ.
B (N. 8.) 5: 84-93, 1956).
Morphology: Sucrose-ammonium agar
most suitable for microscopic study. Pri-
mary and secondary verticils produced,
branches issuing sometimes closely, near the
the sporulating hyphae, forming
tuft-like
spherical to elliptical, 0.6 to 0.8 u.
top of
cluster-like or branches. Spores
Sucrose nitrate agar: Trace of growth.
Glucose-asparagine agar: Growth thin,
moderate, pale olive to pale dark yellow.
Aerial mycelium thin, partially yellowish-
oray.
Nutrient
brown. Aerial mycelium olive-gray to yellow
agar: Growth heavy, deep
to green. Soluble pigment brown.
Potato: Aerial
mycelium yellow-white to yellow-gray. Solu-
Growth heavy, brown.
ble pigment brown.
Milk: Growth brown. Aerial mycelium
scant, yellow-white. Soluble pigment brown.
Gelatin: Liquefaction weak.
Starch: Rapid hydrolysis.
Tyrosinase reaction: None.
Nitrate reduction: Negative.
Cellulose: No growth.
Carbon utilization: Fructose and inositol
utilized. Xylose, rhamnose, sucrose, lactose,
raffinose, and mannitol not utilized.
Habitat: Soil in Japan.
178. Streptomyces omiyaensis Umezawe
et al., 1949 (Umezawa, H., Tazaki, T.,
Okami, Y., and Fukuyama, 8. J. Antibiotics
(Japan) 3: 294-296, 1949).
Morphology: Aerial mycelium
scant branching. Sporophores straight, no
spirals. Spores 1.0 to 1.2 by 2 to 3 u.
Sucrose nitrate agar: Growth thin, trans-
parent, cream-colored to dark. Aerial my-
shows
celium absent, or scant, white. No soluble
pigment.
Nutrient agar: Growth wrinkled, white to
cream-colored. No aerial mycelium. No
soluble pigment. Melanin-negative.
Starch agar: Colorless thin colonies, al-
most all submerged. No aerial mycelium.
No soluble pigment. Hydrolysis.
Gelatin: Growth on surface white. No
soluble pigment. Liquefaction slight in
crateriform.
Potato: Growth white to cream-colored.
No aerial mycelium. No soluble pigment.
Milk: Growth white. Peptonization rapid.
Acid formed.
Antagonistic properties: Produces the
antibiotic chloramphenicol.
Habitat: Soil.
Remarks: Related to S. cacaoz.
179. Streptomyces orientalis Pittenger and
Brigham, 1956 (Pittenger, R. C. and Brig-
ham, R. B. Antibiotics & Chemotherapy 6:
642-647, 1956).
Morphology: Substrate growth made up
of typical prostrate, much-branched myce-
THE ACTINOMYCETES, Vol. II
lium. Aerial mycelium abundant if starch is
used as carbon source. Straight or irregularly
branched sporophores made up of cylindrical
to ovoid spores, 0.7 to 1.0 by 1.4 to 1.8 up.
Sucrose nitrate agar: Growth secant to
moderate, pale cream color. Aerial mycelium
trace of off-white. No pigment or pale yel-
lowish-brown to light brown soluble pigment
may be formed.
Glucose-asparagine agar: Growth moder-
ate to good, cream-colored. Aerial mycelium
pale to cream-colored, powdery. Soluble pig-
ment pale greenish-yellow.
Glycerol malate agar: Growth pale cream
to intense cream-yellow. Aerial mycelium
whitish in color. No soluble pigment. Insolu-
ble malate cleared in agar around growth.
Nutrient agar: Growth cream-colored. Ae-
rial mycelium whitish. No soluble pigment.
Starch agar: Growth moderate, cream-col-
ored to buff to brown. Aerial mycelium
white, becoming pale cream and _ finally
grayish. Soluble pigment cream-yellow, be-
coming pale brown. Hydrolysis limited.
Potato plug: Growth shows slightly rough
surface. Aerial mycelium white. Slight to
moderate amount of brown discoloration of
plug.
Gelatin: Growth flocculent, not forming
intact pellicle. Aerial mycelium scant, white.
No soluble pigment. Liquefaction moderate.
Milk: Heavy wrinkled pellicle, with dull
gray aerial mycelium. No coagulation. Pep-
tonization begins in 11 to 14 days and is
complete in 14 to 21 days. Very dark soluble
pigment obscures litmus color.
Cellulose: Growth good.
Antagonistic properties: Antibiotic vanco-
mycin produced.
temarks: S. orientalis is most closely re-
lated to species intermediate between S.
albus and S. flavus, such as S. alboflavus, S.
fllobisporus, and S. longisporoflavus. S. albo-
gavus cannot utilize cellulose, hydrolyzes
gelatin far less effectively than S. orzentalis,
but attacks starch readily. Milk is feebly
DESCRIPTION OF SPECIES OF STREPTOMYCES 2595
digested by S. alboflavus but rapidly hy-
drolyzed by S. orzentalis. Production of ae-
rial mycelium by the two cultures differs on
several media.
180. Streptomyces paraguayensis — (Al-
meida, 1940) nov. comb. (Almeida, F.
Mycopathologia 2: 201-203, 1940).
Morphology: Thin, ramified mycelial fila-
ments; aerial mycelium consists of thicker
and darker filaments, | u in diameter. Gram-
positive and nonacid-nonalcohol resistant.
Glucose-peptone agar: Growth hard, ad-
hering to the medium; white with dark
center, gradually changing to dark yellow to
almost chocolate.
Nutrient agar: Growth rough, adhering to
the medium; dark gray in color.
Potato: Colonies cerebriform,
growth dry and friable.
Gelatin: Growth on surface. No liquefac-
white;
tion.
Milk: Surface membrane, the milk colored
pink; no peptonization.
Habitat: Thoracic mycetoma; dark heavy
erains.
181. Streptomyces parvullus Waksman and
Gregory, 1954 (Waksman, S. A. and Greg-
ory, F. J. Antibiotics & Chemotherapy 4:
1050-1056, 1954).
Morphology: Sporophores long,
podially branched, twisting into long closed
spirals. Spores spherical, smooth (Pl. I g).
Sucrose nitrate agar: Growth abundant
mono-
with yellow reverse. Aerial mycelium ash-
eray. Soluble pigment yellow.
Glucose-asparagine agar: Growth yellow.
Aerial mycelium abundant, gray. Soluble
pigment yellow.
Nutrient agar: Growth yellowish, covered
with thin white aerial mycelium. Soluble pig-
ment yellow. Melanin-negative.
Potato: Growth orange-colored, covered
with white to gray aerial mycelium. No solu-
ble pigment.
Gelatin: with
Surface pellicle covered
heavy gray aerial mycelium. Liquefaction
slow. Soluble pigment yellow.
Milk: Surface growth heavy, greenish-yel-
low. Aerial mycelium abundant, gray. Solu-
ble pigment brown. No coagulation, very
slow peptonization.
Production of HoS: Negative.
Antagonistic properties: Produces actino-
mycin D.
Habitat: Soil.
Type culture: IMRU 3677.
182. Streptomyces parvus (Xrainsky, 1914)
Waksman and Henrici, 1948 (Krainsky, A.
Centr. Bakteriol. Parasitenk. Abt. II., 41:
685-686, 1914).
Morphology:
branched, or wavy; no true spirals; some
strains, however, produce spirals. Spores
oval, 0:9. to L.d:by 1.2 to-1-8 pu:
Sucrose nitrate agar: Growth yellow, rose
or red. Aerial mycelium light yellow to
white-rose. Soluble pigment rose-colored to
Sporophores straight,
bright yellow.
Calcium malate agar: Small yellow colo-
nies. Light yellow aerial mycelium.
Nutrient agar: Growth yellow. Aerial my-
celum light yellow. Soluble pigment bright
vellow.
Potato: Growth yellow to brown-yellow.
Aerial mycelium white to yellow. Melanin-
negative.
Gelatin: Growth yellow. Soluble pigment
bright yellow. Liquefaction slow.
Milk: No coagulation; rapid peptoniza-
tion.
Starch agar: Growth rose-colored. Aerial
mycelium light gray. Hydrolysis positive.
Cellulose: Growth good, rose-colored. Ae-
rial mycelium yellowish-gray.
Nitrate reduction: Weak.
Production of H.S: Negative.
Antagonistic properties: Produces actino-
mycin.
Habitat: Soil.
Remarks: Ettlinger et al.
(1958) con-
sidered S. parvus as belonging to the S.
griseus series. According to Gause ef al.
(1957), S. parvus is a member of the series
Fradiae.
Type culture: IMRU 3686.
183. Streptomyces pelletiert (Laveran, 1906)
nov. comb. (Laveran, 8. Compt. rend. soc.
biol. 61: 340, 1906).
Morphology: Growth red, smooth, con-
sisting of small, dense, pink colonies. Myce-
lium nonsegmented, branched; hyphae
slender, straight, and not very long. Aerial
hyphae few, straight.
Glucose-asparagine agar: Growth in form
of small, hard, red or purple adherent col-
onies. No soluble pigment.
Glucose agar: Growth poor, in form of
minute, pink colonies.
Glycerol agar: Growth poor, as few moist,
pink colonies.
Nutrient agar: Colonies minute, colorless,
piled up into pale pink masses.
Potato: Growth sparse, yellowish-pink,
irregularly piled up; later, abundant, small,
rounded, pink Aerial mycelium
scant, white.
Blood agar: Colonies at first a few pin-
head, cream-colored; no hemolysis. Later,
dense, button-shaped, with
narrow, fringed margin.
Dorset’s egg medium: Growth abundant,
wrinkled, pink skin with small discrete
colonies at margin; later, surface rough,
masses.
colonies are
mealy, with considerable liquefaction.
Gelatin: Few pink flakes. At first slow,
later almost complete liquefaction.
Milk: Soft curd; gradual peptonization.
Starch: No hydrolysis.
Production of H.S: Negative.
Source: Mycetoma in Nigeria.
Remarks: In the original description of
this culture by Laveran, the organism was
called Micrococcus pelletiert, because no my-
celium was seen, only coccoid bodies. N.
indica was regarded as identical by Pinoy.
N. genesti Froes was described as closely
THE ACTINOMYCETES, Vol. II
allied (Erikson, 1935); the distinction was
founded upon the fact that the red grains
were smaller and much more numerous. A.
africanus is considered as a synonym of this
organism. According to Mariat (1958), S.
pelletiert hydrolyzes gelatin, serum albumin,
casein, and egg albumin; it utilizes urea but
not (NH4).SO, and KNO; as_ nitrogen
sources; it does not utilize xylose, galactose,
maltose, starch, mannitol, or paraffin as car-
bon sources. The species S. africanus is
indistinguishable from S. pelletier?.
184. Streptomyces pentaticus Umezawa
and Tanaka, 1958 (Umezawa, 8. and
Tanaka, Y. J. Antibiotics (Japan) 11A: 26-
29, 1958).
Morphology: Straight sporophores pro-
duce primary and secondary verticils. Spores
can scarcely be observed.
Sucrose nitrate agar: Growth poor, trans-
parent, penetrates deeply into medium. No
aerial mycelium. No soluble pigment.
Glucose-asparagine agar: Growth
less, becoming purplish-pink to dull red-
purple, deep into medium. Aerial mycelium
white, sometimes pink. Soluble pigment
faint brown.
Calcium malate agar: Growth red, irreg-
color-
ular margin. No aerial mycelium. Soluble
pigment faint brown.
Nutrient agar: Growth wet, colorless or
brownish-white. No aerial mycelium. Solu-
ble pigment brown.
Starch Growth
yellow, penetrates deeply into medium. Ae-
rial mycelium white, partially pinkish cot-
agar: colorless or pale
tony colonies. No soluble pigment. Starch
hydrolyzed.
Gelatin: Growth consists of reddish colo-
nies produced on surface. No aerial myce-
lium. Soluble pigment deep brown. Rapid
liquefaction.
Potato: Growth wrinkled, wet, grayish-
brown. No aerial mycelium. Soluble pig-
ment brownish-black.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Milk: Surface ring dull yellow. Coagula-
tion and peptonization.
Antagonistic properties: Produces an anti-
fungal polyenic antibiotic, pentamycin.
Habitat: Soil in Japan.
Remarks: Resembles S.
which differs from the strain producing
rubrireticuli,
pentamycin in the following ways: spirals
are formed; growth on nutrient agar is red;
growth on milk is abundant and red; cellu-
lose and sucrose are utilized.
185. Streptomyces phaeochromogenes
(Conn, 1917) Waksman and Henrici, 1948
(Gonn: HH. J. (N. Y: Agr.. Expt: Sta. Tech.
Bulle 60.1917).
This culture has been studied by Conn
(1917), Waksman (1919), Jensen (1931),
Krassilnikov (1949), Kutzner (1956), and
Ettlinger et al. (1958).
Morphology: Sporophores form narrow,
open, elongated, sinistrorse spirals (Conn,
Waksman, Jensen, Krassilnikov). Kutzner
(1956) examined 25 strains belonging to this
species; only five of them produced spirals.
Ettlnger et al. (1958) could not find any
spirals on any of the strains obtained from
various culture collections. Spores spherical
to short rods; surface smooth (Pl. I d).
Sucrose nitrate agar: Growth brown to
almost black. Aerial mycelium abundant,
white with brownish shade. Soluble pigment
brown to dark brown.
buff to
brown. Aerial mycelium white. Soluble pig-
Calcium malate agar: Growth
ment brown.
Nutrient
later turning nearly black. Aerial mycelium
agar: Growth gray to brown,
white to gray, often absent. Soluble pigment
deep red-brown.
Starch agar: Growth brown. Hydrolysis
medium.
Potato: Growth brown to almost black.
No aerial mycelium. Soluble pigment dark
brown to black.
Gelatin: Surface
growth abundant,
spreading, cream-colored, becoming brown.
Liquefaction slow. Soluble pigment brown.
Milk: Dark, almost black ring:
tion with slow peptonization.
Nitrate: Reduction limited.
Production of H.S: Positive.
Temperature: Optimum 25°C.
Antagonistic properties: Strong.
Habitat: Soil.
Type culture: IMRU 3338.
coagula-
186. Streptomyces phaeopurpureus Shi-
nobu, 1957 (Shinobu, R. Mem. Osaka Univ.,
B. Nat. Sci. 6: 638-67, 1957).
Morphology: Substrate mycelium mono-
podial, 0.4 to 0.6 uw in diameter; no frag-
mentation. Aerial mycelium straight, usu-
ally short. Spores spherical to elliptical, 0.6
to 0.8 uw; rarely 1 yu.
Sucrose nitrate agar: Growth good, brown
to dark red. No aerial mycelium. Soluble
pigment brown.
Glycerol malate
orange to purple. Aerial mycelium powdery,
vellowish-gray to pinkish-gray. Soluble pig-
ment red-purple to brown-purple.
Glucose-asparagine agar: Growth moder-
agar: Growth good,
ate, orange to red-brown. Aerial mycelium
moderate, in patches, pinkish-gray. Soluble
pigment reddish-orange to reddish-brown.
Nutrient agar: Growth good, deep brown.
No aerial mycelium. Soluble pigment brown
to deep reddish-brown.
Potato plug: Growth wrinkled, reddish to
yellowish-brown. Aerial mycelium absent or
scant, light brownish-gray. Plug colored
brown.
Milk: Growth in form of deep brown ring.
Soluble pigment brown. No coagulation;
peptonization uncertain.
Tyrosinase reaction: Positive.
Gelatin: Liquefaction fairly strong.
Diastase: Weak.
Cellulose: Negative.
Nitrate reduction: Negative.
Carbon sources: Utilizes xylose, rhamnose,
258 THE ACTINOMYCETES, Vol. II
fructose, sucrose, lactose, raffnose, mannitol,
and inositol.
Habitat: Soil in Japan.
187. Streptomyces phaeoviridis Shinobu,
1957 (Shinobu, R. Mem. Osaka Univ., B.
Nat. Sci. 6: 67-70, 1957).
Morphology: Growth monopodial, hyphae
0.4 to 0.6 uw in diameter, no fragmentation.
Aerial mycelium short; monopodial branch-
ing; some spirals, sinistrorse, 1 to 8 turns.
Spores elliptical, 0.6 to 0.8 uw.
Sucrose nitrate agar: Growth pale yellow
to dark brown. Aerial mycelium scant,
brownish-white. Soluble pigment yellow to
brown to dark blue.
Malate-glycerol agar: Growth yellowish-
brown to dark brown. Aerial mycelium scant,
white to brownish-white. Soluble pigment
brown to dark brown with blue tinge.
Glucose-asparagine agar: Growth thin
brown to yellow-orange; aerial mycelium
scant, white to brownish-white. Soluble
pigment pale brown to yellow-orange.
Nutrient agar: Growth thin, yellow-orange
to brown. No aerial mycelium. Soluble pig-
ment brown to dark red with purple tinge.
Potato: Growth poor, pale brown. No
aerial mycelium. Soluble pigment uncertain,
probably pale brown.
Milk: Growth good, pale yellow, partially
blue, sometimes grayish-green. Soluble pig-
ment absent or pale orange. Coagulation
and peptonization.
Tyrosinase reaction: Negative.
Gelatin: Liquefaction variable.
Diastase: Strong.
Nitrate reduction: Negative.
Cellulose: No growth.
Carbon sources: Utilizes xylose, rhamnose,
sucrose, fructose, raffinose, and mannitol;
lactose and inositol uncertain.
Habitat: Soil in Japan.
I88. Streptomyces pilosus Ettlinger et al.,
1958 (Ettlinger, L., Corbaz, R., and Hitter,
R. Arch. Mikrobiol. 31: 347, 1958).
Morphology: Sporophores monopodially
branched, with long, regular, open spirals.
Spores covered with fine long hair.
Glycerol nitrate agar: Growth yellow to
yellow-brown. Aerial mycelium powdery,
chalk-white to gray-blue.
Glucose-asparagine agar: Growth light
yellow. Aerial mycelium powdery, white to
ash-gray.
Calcium malate agar: Growth white-yel-
low to yellow-brown. Aerial mycelium white-
yellow to white-gray.
Starch agar: Growth yellow-brown to red-
yellow. Aerial mycelium scant, white to
ash-gray. Limited hydrolysis.
Gelatin: Growth yellow-brown. Aerial
mycelium powdery, chalk-white. Liquefac-
tion slow. Soluble pigment dark brown.
Potato: Growth golden yellow. Aerial
mycelium ash-gray. Soluble pigment dark
brown.
Milk: Surface growth white-gray to gray-
ish-blue. Aerial mycelium ash-gray. No co-
agulation; peptonization weak.
Antagonistic properties: Positive.
Habitat: Soil from Rome, Italy.
189. Streptomyces platensis Pittenger and
Gottlieb, 1954 (Pittenger, R. C. and Gott-
heb, D. Brit. Pat. 713,795, August’ 18,
1954*).
Morphology: Aerial mycelium forms loose
to tight spirals on its sporophores. Spores
ovoid, 0.7 to 0.9 by 0.8 to 1.2 yu.
Sucrose nitrate agar: Substrate growth
deep olive, reverse becoming dark olive.
Aerial mycelium pale smoke-gray with tufts
of white; areas of black pigmented aerial
growth may also be found, giving effect of
a mosaic.
Glucose - asparagine agar: Substrate
erowth ochraceous-buff becoming tawny.
Aerial mycelium white becoming grayish-
olive to almost black. Soluble pigment ab-
sent or shght, brown.
* Supplemented by personal communication.
DESCRIPTION OF SPECIES OF STREPTOMYCES 259
Calcium malate agar: Growth ochraceous-
salmon, becoming cinnamon-buff. Aerial
mycelium quaker-drab with areas of black
and white. Soluble pigment slight, greenish-
yellow.
Nutrient agar: Poor substrate growth,
cream-yellow becoming buff to drab. No
aerial mycelium. Slight soluble brown pig-
ment.
Starch agar: Growth cream- to buff-col-
ored. Aerial mycelium white, becoming
mouse-gray with patches of black. Slow
hydrolysis.
Potato: Excellent growth. Aerial myce-
lium white to pale mouse-gray. Soluble pig-
ment brown.
Gelatin: Very slow liquefaction. Melanin-
negative.
Milk: Growth secant, forming partial ring
at surface. No coagulation or peptonization.
Blood: Hemolysis.
Cellulose: Growth slight. Aerial mycelium
gray to black.
Nitrate: Reduction to nitrite, especially
with starch as source of carbon.
Carbon utilization: Starch, malic acid,
inositol, sodium succinate, sodium citrate,
sorbitol, mannitol, maltose, arabinose, lac-
tose, galactose, fructose well utilized. Dul-
citol, raffinose, cellulose, sodium formate,
sodium tartrate, xylose poorly utilized.
Asparagine, rhamnose, o0-cresol, m-cresol,
sodium acetate, inulin, sodium. salicylate
not utilized.
Antagonistic properties: Produces oxytet-
racycline.
Remarks: Tresner and Backus (1956) con-
sidered this organism as a variant of S.
hygroscopicus rather than a separate species.
Ettlinger et al. (1958) came to similar con-
clusions.
190. Streptomyces pluricolor (Berestnew,
1897 emend. Krassilnikov, 1941) Waksman
(Krassilnikov, N. <A. Actinomycetales.
Izvest. Akad. Nauk. SSSR, Moskau, p. 17,
1949).
Morphology: Sporophores produce nu-
merous spirals, with 3 to 5 turns (sinistrorse).
Spores oval, 0.9 by 0.7 u.
Synthetic agar: Growth at first pigmented
vellow-red, later becoming blue to blue-
green. Aerial mycelium white-gray. The blue
pigment dissolves into the medium.
Nutrient agar: Soluble pigment greenish,
fluorescent.
Potato: Growth and soluble pigment
sharp blue.
Gelatin: Liquefaction rapid.
Milk: Peptonization positive; no coagula-
tion.
Starch: Hydrolysis.
Cellulose: No growth.
Nutrient broth: Soluble pigment green,
fluorescent.
Sucrose: Inversion.
Antagonistic properties: None.
Habitat: Soil.
temarks: Closely related to S. violace-
oruber. A. pluricolor diffundens Berestnew is
considered by Krassilnikov as a synonym.
191. Streptomyces pluricolorescens Okami,
Y. and Umezawa, H.* n. sp.
Morphology: Aerial hyphae not flexuous,
few branches; no spirals.
Glycerol nitrate agar: Growth at first
and yellowish, then yellowish-
brown with reddish tone. Aerial mycelium
white to olive or pinkish. Soluble pigment.
shghtly yellowish-brown or light wine-color
with aging.
Glucose—asparagine agar: Growth at first
and yellowish, then yellowish-
brown with reddish tone. Aerial mycelium
white to olive or pinkish. Soluble reddish-
purple pigment occasionally produced.
Calcium malate agar: Same as on glycerol
nitrate agar.
colorless
colorless
|Q70
Nutrient agar: 37°C. Colorless or slight
vellowish-brown growth with fine wrinkles.
Aerial mycelium white. Soluble pigment ab-
sent at first, later a brown pigment appears.
* Personal communication from Okami.
260
Potato plug: Growth colorless, then
slightly yellowish or brownish. Aerial myce-
lium white to olive-colored. Color of plug
unchanged. Melanin-negative.
Gelatin: 18-20°C. Growth colorless. No
aerial mycelium. Soluble pigment. slightly
yellowish-brown. Gelatin liquefied.
Milk: 37°C. Colorless to slight yellowish
growth. No aerial mycelium. Coagulation
with acid reaction, then peptonization.
Starch: Hydrolysis weak to medium.
Carbon utilization: Good growth with
arabinose, dextrin, fructose, galactose, glu-
cose, glycerol, maltose, mannitol, mannose,
raffinose, rhamnose, salicin, sorbose, starch,
sucrose, xylose, and sodium succinate. Scant
growth with esculin, inositol, lactose, sor-
bitol, sodium acetate, and citrate.
Antagonistic properties: Produces anti-
tumor substances pluramycin A and B
(Maeda et al., 1956).
temarks: This culture is said to be re-
lated to S. vinaceus, but it does not produce
blue-red pigment in reverse of growth on
nutrient agar.
192. Streptomyces poolensis (Taubenhaus,
1918) Waksman (Taubenhaus, J. J. J. Agr.
Xesearch 13: 446, 1918).
Morphelogy: Sporophores straight. Spores
oval to elliptical.
Sucrose nitrate agar: Growth thin, color-
less. Aerial mycelium white to gray.
Glucose-asparagine agar: Growth abun-
dant, glossy, light brown.
Nutrient agar: Growth translucent, yel-
lowish to brown. Soluble pigment brown.
Potato: Growth thin, reddish-brown. Sol-
uble pigment purplish.
Gelatin: Liquefaction, with small, brown-
ish flakes in fluid.
Milk: Brownish ring. Coagulation and
peptonization.
Starch: Growth restricted, cream-colored.
No hydrolysis.
Nitrate reduction: Positive.
Antagonistic properties: Positive.
THE ACTINOMYCETES, Vol. II
Habitat: Sweet-potato disease known as
pOxs ”
193. Streptomyces praecox (Millard and
Burr, 1926) Waksman and Henrici, 1948
(Millard, W. A. and Burr, 8. Ann. Appl.
Biol. 13: 580, 1926).
Morphology: Sporophores produce short,
open spirals. Spores spherical or oval, 0.8
uw in diameter.
Sucrose nitrate agar: Growth thin, color-
less. Aerial mycelium gray to olive-buff. On
continued cultivation, aerial mycelium tends
to become white.
Nutrient agar:
mycelium white.
Starch media: Growth thin, cream-col-
ored. Aerial mycelium white with greenish
tinge. Hydrolysis positive.
Potato: Growth lichenoid, cream-colored
to light brown. Aerial mycelium white to
olive-buff. Soluble pigment olive-buff to
drab. On continued cultivation, no soluble
pigment produced.
Gelatin: Growth good. Aerial mycelium
white. Liquefaction medium. Melanin-nega-
tive.
Milk: Surface growth cream-colored, in
Growth colorless. Aerial
form of ring. Aerial mycelium white. Coagu-
lation slow; peptonization rapid.
Nitrate: Reduction variable.
Cellulose: Good growth, colorless. Aerial
mycelium dark gray.
Tyrosinase reaction: Negative.
Production of H.S: Negative.
Temperature: Grows well at 37.5°C
Odor: Very strong.
Antagonistic properties: Represses growth
of S. scabies.
Habitat: IKnoblike scab of potatoes.
temarks: According to Ettlinger et al.
(1958), this organism belongs to the S.
griseus series. Hoffmann (1958) described a
culture of S. praecox that produced a light
to dark gray aerial mycelium and many
spirals; nonchromogenic.
Type culture: IMRU 3374.
DESCRIPTION OF SPECIES OF STREPTOMYCES
194. Streptomyces praefecundus (Millard
and Burr, 1926) Waksman (Millard, W. A.
and Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight, fre-
quently forming brushes. Spores spherical
to oval, 0.8 by 0.85 wu.
Sucrose nitrate agar: Growth good, cream-
colored. Aerial mycelium cottony, olive-
buff. Soluble pigment cream-colored.
Nutrient potato agar: Growth lichenoid,
gray. Aerial mycelium smooth, white to
yellowish. Soluble pigment golden brown.
Potato: Growth good, wrinkled. Aerial
mycelium white to yellowish to olive-buff.
Soluble pigment gray to brown.
Gelatin: Surface growth good. Aerial my-
celium white. Soluble pigment light pink to
dark golden brown. Liquefaction rapid.
Milk: Surface growth good. Aerial myce-
lium scant, white. Coagulation and peptoni-
zation.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Temperature: Grows well at 37.5°C.
Habitat: Potato scab and soil.
195. Streptomyces prasinophilus Ettlinger
et al., 1958 (Ettlinger, L., Corbaz, R., and
Hiitter, R. Arch. Mikrobiol. 31: 345, 1958).
Morphology: Sporophores monopodially
branched, long, straight, with open spirals,
usually 1 to 8 coils. Spores covered with
long, fine hair (Pl. II, Ix).
Glycerol nitrate agar: Growth red or red-
brown. Aerial mycelium leek-green
white spots. Soluble pigment pink.
Glucose-asparagine agar: Growth brick-
red. Aerial mycelium white to leek-green.
Soluble pigment brick-red.
Glycerol malate agar: Growth brick-red.
Aerial mycelium leek-green. Soluble pig-
ment pink.
Glucose-peptone agar: Growth brick-red.
Starch-KNO; agar: Growth pink. Aerial
mycelium white to leek-green. Soluble pig-
ment light pink. Good hydrolysis.
Gelatin: Bottom flakes red to yellowish,
with
261
later brick-red. Slow liquefaction. No soluble
pigment. Melanin-negative.
Potato: Growth slow, flesh-red. No solu-
ble pigment.
Milk: Strong coagulation and peptoniza-
tion.
Antagonistic properties: Weak activity
against gram-positive bacteria.
Habitat: Soil in Mallorca, Spain.
196. Streptomyces prasinus Ettlinger et al.,
1958 (Ettlinger, L., Corbaz, R., and Hitter,
R. Arch. Mikrobiol. 31: 343, 1958).
Morphology: Sporophores monopodially
branched, long, straight, with open spirals,
forming | te 2 coils. Spores covered with
short spines (PI. IT, Ix).
Glycerol nitrate agar: Growth colorless.
Aerial mycelium grass-green, later dark
green.
Glucose-asparagine agar: Growth whitish-
vellow.
Glycerol malate agar: Growth copper-red.
Aerial mycelium velvety, leek-green.
Gelatin: Growth limited, whitish-yellow.
No soluble pigment. Liquefaction positive.
Starch agar: Growth reddish-brown.
Aerial mycelium leek-green. Strong hy-
drolysis.
Potato: Growth limited, lght brown.
Aerial mycelium leek-green. Melanin-nega-
tive.
Milk: Heavy pellicle. Aerial mycelium
whitish-gray to greenish-gray. No coagula-
tion; slow peptonization.
Antagonistic properties: None.
Habitat: Soils in Mallorea and Belgian
Congo.
197. Streptomyces pseudogriseolus Okami
et al., 1955 (Okami, Y., Utahara, R., Oyagi,
H., Nakamura, S., and Umezawa, H. J.
Antibiotics (Japan) 8A: 126-131, 1955).
Morphology: Sporophores produce nu-
merous closed spirals. Spores oval to cylin-
drical, 0.8 to 1:2 by 1:0 to: 1:5 py.
Glycerol nitrate agar: Growth colorless to
262
grayish-buff. Aerial mycelium grayish-buff,
powdery. No soluble pigment.
Nutrient agar: Growth colorless. Aerial
mycelium white, thin. Soluble pigment ab-
sent or shght, brown.
Potato: Growth colorless to slightly yel-
lowish, elevated, wrinkled. Aerial mycelium
white, cottony to velvety. No soluble pig-
ment.
Milk: Surface growth orange. Aerial my-
celium velvety, white. Coagulation and pep-
tonization completed in 25 to 30 days.
Gelatin: Growth yellowish-brown. Aerial
mycelium white to grayish. Soluble pigment
brownish. Liquefaction weak to medium.
Melanin-negative.
Starch: Hydrolysis strong.
Carbon utilization: Utilizes arabinose,
dextrin, 7zso-dulcitol, fructose, galactose,
glucose, glycerol, inositol, lactose, maltose,
mannitol, mannose, rhamnose, — salicin,
starch, sucrose, sodium acetate, sodium
citrate, and sodium succinate. Does not
utilize esculin, raffinose, sorbitol, or sorbose
(inulin).
Antagonistic properties: Produces xantho-
mycin-like substance.
Habitat: Isolated from soil in Japan.
Remarks: This culture resembles S. gr7-
seolus, but it differs in spiral formation and
growth on potato and milk media. These
differences may not be enough to warrant
establishing a new species, but the produc-
tion of xanthomycin is a property not found
in the culture liquid of S. griseolus.
Type culture: ATCC 12,770.
198. Streptomyces purpureefuscus Yama-
guchi and Saburi, 1955 (Yamaguchi, T. and
Saburi, Y. J. Gen. Appl. Microbiol. 1: 201-
235, 1955).
Morphology: Aerial hyphae and
straight; on synthetic and starch agars, they
long
show a tuft-forming tendency at the margin.
Spirals not produced. Spores cylindrical,
1.1 to 2.2 by 0.7 to 1.1 py.
Sucrose nitrate agar: Growth thin, color-
THE ACTINOMYCETES, Vol. II
less, later becoming purple to dark purple;
this property may be lost after repeated
transter, in which case growth remains white
to hght purple. Aerial mycelium powdery,
white, later smoke-gray. Soluble pigment
faint purple.
Glycerol malate agar: Growth brownish
to purplish-brown. Aerial mycelium white,
smoke-gray to light grayish-olive. Soluble
pigment light brownish-purple.
Nutrient agar: Growth wrinkled, colorless.
Aerial mycelium absent, or scant, white.
Soluble pigment changes from reddish-brown
to dark vinaceous-brown.
Starch agar: Growth yellowish-brown to
dark olive-buff; sometimes with hygroscopic,
black patches. Aerial mycelium velvety, at
first white, later olive-gray. Usually no sol-
uble pigment is produced, but sometimes
faint pinkish-purple is seen. Strong hy-
drolysis.
Potato: Growth vigorous, finely wrinkled,
at first purplish-brown or yellowish-brown,
later becoming black. Aerial mycelum abun-
dant, powdery, grayish. Soluble pigment
purplish.
Gelatin: Growth dark brownish-gray.
Aerial mycelium coarse, powdery, grayish-
white. Soluble pigments yellowish-brown to
brown and a more diffusible yellowish-green.
Strong liquefaction.
Milk: Growth at first dull reddish-brown,
later purplish-brown. Soluble pigment faint
purplish and more diffusible faint yellowish.
Coagulation and peptonization.
Cellulose: No growth.
Carbon utilization: pD-xylose, L-arabinose,
b-galactose, sucrose, lactose, raffinose, so-
dium succinate readily utilized. L-rhamnose,
inulin, mannitol, sorbitol, inositol, acetate,
and citrate not utilized.
Antagonistic properties: Active against
gram-positive and acid-fast bacteria; pos-
sesses antitrichomonal activity.
Remarks: Related to S.
cylindrosporus, and S. purpureochromogenes.
vinaceus, S.
DESCRIPTION OF SPECIES OF STREPTOMYCES
199. Streptomyces purpurascens Linden-
bein, 1952 (Lindenbein, W. Arch. Mikrobiol.
17: 361-383, 1952).
Morphology: Sporophores long, straight,
with open and closed spirals, 2 to 5 turns as
side branches. Spores covered with long
spines (Pl. Ib). A detailed electron micro-
scope study of this organism has been made
by Petras (1959).
Glycerol nitrate agar:
red to purple. Aerial mycelium cottony,
white to purplish. Soluble pigment brown-
Growth carmine-
red.
Glucose-asparagine agar: Growth car-
mine-red to purple. Aerial mycelium white
to pinkish. Soluble pigment orange to car-
mine-red.
Glycerol malate agar: Growth carmine-
red. Aerial mycelium white. Soluble pigment
orange to brick-red.
Nutrient agar: Growth light brown, with
dark brown reverse. Aerial mycelium white.
Soluble pigment dark brown. Melanin-
positive.
Glucose-peptone agar: Growth lichenoid,
red to red-brown. Aerial mycelium white.
Soluble pigment light brown.
Starch media: Growth light carmine to
vellow-red. Aerial mycelium white. No sol-
uble pigment. Hydrolysis strong.
Potato: Growth reddish.
Aerial mycelium white to gray. No soluble
pigment. (Kutzner (1956) observed on six
strains a gray to black pigment on potato
plug.)
Gelatin: Growth light brown. Aerial my-
celium white. Soluble pigment red-brown.
Liquefaction medium.
Milk: Growth red to dark brown. Aerial
mycelium white. No proteolysis.
Cellulose: Growth white to red.
Production of HoS: Positive.
Antagonistic properties: Produces rhodo-
mycin.
Remarks: On continued growth on syn-
thetic media, the culture may the
brownish to
lose
263
property to produce the typical pigment. It
can be regained, however, by growth on
organic media. This organism is considered
by Corbaz et al. (1957) as a synonym of S.
bobiliae, except that the latter lost the prop-
erty of producing aerial mycelium or spores.
Lindenbein (1952) and Frommer (1959) ob-
tained colorless mutants from S. purpuras-
cens.
Type culture: IMRU 3660.
200. Streptomyces purpureochromogenes
(Waksman and Curtis, 1916) Waksman and
Henrici, 1948 (Waksman, S. A. and Curtis,
R. BE: Soil Sar Ts 23. 1916: S132 71919).
Morphology: Long sporophores produce
few imperfect spirals. Spores spherical, 0.75
to 1.0 uw in diameter.
Sucrose nitrate agar:
smooth, gray, becoming brown with purplish
tinge; center raised, margin yellow. Aerial
mycelium dark brown to dark gray.
Glucose-asparagine agar: Growth abun-
dant, gray, becoming brown to dark brown.
Nutrient agar: Growth gray to brownish,
becoming dark brown, almost black. Soluble
Growth restricted,
pigment dark brown. Melanin-positive.
Potato: Growth orange to orange-red.
Gelatin: Surface growth slow, brownish.
Liquefaction slow.
Milk: Dark brown ring. Coagulation and
slow peptonization.
Starch media: Colonies small, dark brown.
Slight hydrolysis.
Cellulose: Moderate growth.
Sucrose: Inversion.
Nitrate reduction: Negative.
Production of H.S: Negative.
Temperature: Optimum 25°C.
Antagonistic properties: Active against
various bacteria.
Habitat: Soil.
Type culture: IMRU 3343.
201. Streptomyces putrificus (Nikolaieva,
1915) Waksman (Nikolaieva, E. Arch. Biol.
Nauk. 18: 229, 1914).
264
Morphology: Sporophores spiral-shaped.
Spores spherical.
Nutrient agar: Growth colorless to gray-
ish. Aerial mycelium white. No soluble pig-
ment.
Potato: Growth folded, sulfur-yellow.
Aerial mycelium chalk-white. No soluble
pigment. Melanin-negative.
Milk: Pellicle heavy. Aerial mycelium
white. Peptonization gradual without pre-
vious coagulation.
LoefHer’s serum: Growth yellow. No aerial
mycelium. Serum liquefied and colored yel-
lowish-brown.
Odor: Strong, putrefactive.
Habitat: Spring water.
Remarks: Decomposes proteins ener-
getically, with the formation of bad-smelling
products (HS, NH;). Morphological prop-
erties given by Krassilnikov (1949), who
considers this organism as a member of the
A. albus group.
202. Streptomyces pyridomyceticus Okami
et al., 1957 (Okami, Y., Maeda, K., and
Umezawa, H. J. Antibiotics (Japan) 7A:
55-56, 1954; 10A: 172, 1957).
Morphology: Sporophores form flexible,
open spirals. Spores of irregular size.
Glycerol nitrate agar: Growth colorless
to dark. Aerial mycelium thin, white, some-
times gray to brownish-gray. No soluble
pigment.
Nutrient agar: Growth colorless. Aerial
mycelium absent, or scant, white. No soluble
pigment.
Potato: Growth wrinkled, dark yellowish-
brown. Aerial mycelium absent, or later
white. No soluble pigment.
Gelatin: Growth colorless. Aerial myce-
lium white, sometimes grayish. No soluble
pigment. No liquefaction.
Starch: Hydrolysis.
Milk: Growth yellowish, in the form. of
surface ring. No aerial mycelium. Coagula-
tion and peptonization absent or very slow.
Blood: No hemolysis.
THE ACTINOMYCETES, Vol. II
Nitrate reduction: Negative.
Carbon utilization: Utilizes
dextrin, fructose, galactose, glucose, glycerol,
maltose, xylose, and sucrose. Does not utilize
arabinose,
dulcitol, esculin, inulin, lactose, mannose,
raffinose, rhamnose, salicin, sorbitol, sodium
citrate, sodium acetate, and sodium suc-
cinate.
Habitat: Soil in Japan.
Antagonistic properties: Produces anti-
biotic pyridomycin.
temarks: Isolated by means of chlortetra-
cycline-containing agar. Related to S. cacaoz
and S. flocculus, as well as to S. hygroscopicus.
Above description was first given under the
name S. albzdofuscus. It was later found that
this name was preempted by Neukirch and
Berestnew, and was, therefore, changed to
S. pyridomyceticus.
203. Streptomyces rameus Okami et al.,
1959) (Okami, Y., Suzuki, M., Takita, T.,
Ohi, IK., and Umezawa, H. J. Antibiotics
(Japan) 12A: 257-262, 1959).
Morphology: Aerial mycelium forms in-
complete spirals or loops or hooks. Spores
oval to oblong.
Glycerol nitrate agar: Growth yellow.
Aerial mycelium white. Soluble pigment
absent or yellowish.
Glucose-asparagine agar: Growth yellow-
ish. Aerial mycelium white. Soluble pig-
ment absent or yellowish.
Calcium malate agar: Growth colorless to
yellowish. Aerial mycelium off-white. No
soluble pigment.
Nutrient agar: Growth colorless to brown-
ish. Aerial mycelium white. Soluble pigment
absent or shght brownish.
Starch agar: Growth colorless to yellowish.
Aerial mycelium white. Soluble pigment
yellowish. Hydrolysis weak to medium,
Potato: Growth brownish. Aerial myce-
lium white. Soluble pigment brown to black.
Gelatin: Growth brownish. Aerial myce-
lium white. Soluble pigment brown. Liquetac-
tion doubtful.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Milk: Coagulation weak; peptonization
doubtful.
Nitrate reduction: Negative.
Carbon sources: Utilizes arabinose, dex-
trin, fructose, galactose, glucose, glycerol,
inositol, maltose, mannitol, mannose, rafh-
nose, starch, and sucrose. Lactose and xylose
gave less response. Poor growth on inulin,
rhamnose, salicin, sorbitol, sorbose, sodium
acetate, and sodium citrate.
Antagonistic properties: Produces strepto-
mycin.
ftemarks: Related to S. alboflavus, S.
xanthophaeus, and S. orventalis.
204. Streptomyces ramnait Bhuiyan and
Ahmad, 1956 (Bhuiyan, A. M. and Ahmad,
K. Ann. Biochem. Exptl. Med. India 16:
101-104, 1955).
Morphology: Open spirals, with 2 or 3
turns. Spores spherical, 0.8 « in diameter.
nitrate Growth whitish.
Aerial mycelium white to pale rose. No
soluble pigment.
Glucose-asparagine agar:
Sucrose
agar:
Growth color-
less to pale rose. Aerial mycelium white,
later pale rose.
Calcium malate agar: Growth smooth,
cream-colored. Aerial mycelium white. Me-
dium becomes clear.
Nutrient agar: Growth cream-colored, be-
coming light brown. Aerial mycelium pow-
dery, white. Soluble pigment shght brown
coloration. Melanin-negative.
Starch agar: Growth
yellowish brown. No aerial mycelium. No
soluble pigment. Hydrolysis rapid.
cream-colored to
Potato: Growth abundant, cream-colored.
Aerial mycelium white, turning pale rose.
Potato nutrient agar: Growth rapid, color-
less to cream-colored. Aerial mycelium deep
rose. Soluble pigment deep reddish-brown
to almost red.
Gelatin: Growth cream-colored. No aerial
mycelium. Soluble pigment absent or light
brown. Liquefaction medium.
Milk: Growth cream-colored. No aerial
mycelium. Coagulation, followed by
tonization. Reaction acid.
Nitrate reduction: Positive.
Cellulose: Growth good.
pep-
Optimum temperature: 37°C.
Antagonistic properties: Produces
biotic ramnacin.
anti-
205. Streptomyces ramulosus Ettlinger et
al., 1958 (Ettlinger, L., Giumann, E., Hiit-
ter, R., IKeller-Schierlein, W., Kradolfer,
F., Neipp, L., Prelog, V., and Zahner, H.
Helv. Chim. Acta 41: 216-219, 1958).
Morphology: Sporophores monopodially
branched, straight with many side branches.
Spores smooth (PI. IT h).
Glycerol nitrate agar: Growth at first
‘armine-red, later turning greenish-brown.
Aerial mycelium ash-gray with greenish
tinge. Substrate pigmented carmine-red.
Glucose-asparagine agar: Growth yellow-
ish-red. Aerial mycelium gray. Substrate
carmine-red.
Calcium malate agar: Growth hght vellow.
Aerial mycelium chalky white to gray with
greenish tinge.
Glucose-peptone agar: Growth yellowish-
red, partly greenish to greenish-black. Aerial
mycelium powdery, Substrate
greenish to brownish-black.
Starch agar: Growth light yellow. Sub-
strate ight carmine. Gradual hydrolysis.
Potato: Growth yellowish-red. Aerial my-
celium chalk-white to ash-gray. Substrate
carmine-red.
Gelatin: Growth light yellow. Soluble pig-
ment light brown. No liquefaction.
Milk: Light yellow pellicle. No aerial
mycelium. Coagulation limited; peptoniza-
tion good.
ash-gray.
Carbon. utilization: Glucose, L-xylose, p-
inulin, well
utilized. Does not utilize raffinose, L-arabi-
p-mannitol, Question-
able utilization of L-rhamnose, salicin.
fructose, sucrose, p-sorbitol
mesoinositol.
nose,
Antagonistic properties: Produces anti-
biotic acetomycin, active against gram-posi-
266
tive and gram-negative bacteria, as well as
against trichomonads and amoebae.
206. Streptomyces resistomycificus Linden-
bein, 1952 (Lindenbien, W. Arch. Mikro-
biol. 17: 361-883, 1952).
Morphology: Sporophores
curling tips. Spores short, oval.
long, with
Glycerol nitrate agar: Growth yellow-
brown to dark brown. Aerial mycelium ash-
gray. Soluble pigment red-brown.
Glucose-asparagine agar: Growth yellow-
brown. Aerial mycelium ash-gray. Soluble
pigment yellow-brown.
Glycerol malate agar: Growth dark brown.
Aerial mycelium ash-gray to red-gray.
Soluble pigment gray to dark brown.
Nutrient agar: Growth dark brown.
Aerial mycelium absent or lead-gray. Soluble
pigment dark brown. Melanin-positive.
Glucose-peptone agar: Growth dark
brown. Aerial mycelium white. Soluble pig-
ment reddish to dark brown.
Starch agar: Growth light yellow to red-
dish-brown. Aerial mycelium gray-white,
later red-gray. Soluble pigment lacking or
reddish-brown. Hydrolysis strong.
Potato: Growth brownish-black. Aerial
mycelium reddish-white. Soluble pigment
dark brown.
Gelatin: Growth dark brown. Aerial my-
celium white-gray. Soluble pigment chest-
nut-brown. Good liquefaction.
Milk: Growth dark brown. Aerial myce-
lium white, later yellowish-red. Soluble pig-
ment dark brown. Peptonization none or
slight.
Cellulose: No growth.
Antagonistic properties: Produces resisto-
mycin, which is active against gram-positive
bacteria.
Remarks: Gause et al. (1957) have de-
scribed certain closely related forms, such as
A. griseorubiginosus with a variety spiralis,
and A. variabilis with a variety roseolus.
Type culture: IMRU_ 3658.
THE ACTINOMYCETES, Vol. II
207. Streptomyces reticuli (Waksman and
Curtis, 1916; Waksman, 1919) Waksman and
Henrici, 1948 (Waksman, 8. A. and Curtis,
R. E. Soil Sci. 1: 118, 1916; Waksman, 8. A.
Soil Sci. 8: 143, 1919).
Morphology: Aerial mycelium gives rise
to simple verticils. Sporophores straight or
spiral-shaped (sinistrorse) on different media.
Spores spherical or oval, smooth, 1.0 to 1.4
uw in diameter (PI. I a).
Sucrose nitrate agar: Growth colorless,
with yellowish tinge, becoming brownish.
Aerial mycelium thin, cottony, white to ash-
gray. No soluble pigment.
Glycerol malate agar: Growth colorless.
Aerial mycelium yellowish. No soluble pig-
ment.
Nutrient agar: Growth wrinkled, gray,
becoming brownish. No aerial mycelium.
Soluble pigment dark brown.
Potato: Growth gray, with black center.
Aerial mycelium ash-gray. Soluble pigment
black.
Gelatin: Growth gray to brown. Aerial
mycelium white. Soluble pigment faint
brown to dark brown. Good liquefaction.
Milk: Coagulation rapid; peptonization
slow.
Starch: Growth brownish-gray. Hydrol-
ysis.
Cellulose: Scant growth.
Nitrate reduction: Positive.
Production of H.S: Positive.
Invertase: Positive.
Temperature: Optimum 25°C.
properties:
produce neomycin or a neomycin-like sub-
stance. Some strains reduce double bonds in
Antagonistic Some strains
certain steroids.
Habitat: Soil.
Remarks: According to Ettlinger et al.
(1958), the verticils are both primary and
secondary; no spirals were observed. They
also report the species to be melanin-nega-
tive. One wonders whether they had a typi-
cal culture. This culture was later found to
DESCRIPTION OF SPECIES OF STREPTOMYCES
be identical with S. abzkoensum. Sakagami
et al. (1958) described a variety latumcidicus
that produced no aerial mycelium on most
media and formed the antibiotic latumeidin.
Type culture: IMRU 3344.
208. Streptomyces rimosus Sobin et al.,
1950 (Sobin, B. A., Finlay, A. C., and Kane,
J. H. U.S. 2,516,080, July 18, 1950; see also
Kochi, M., eé al. Proc. Natl. Acad. Sci. U.S.
38: 383-391, 1952).
Morphology: Sporophores long, usually
straight, occasionally open or closed spirals
depending on composition of medium. Speres
cylindrical, 0.6 to 0.7 by 0.8 to 1.4 ». A
microscopic study of S. rzmosus (strain 3558)
grown on yeast extract-glucose agar, after
14 days incubation, revealed the following:
Aerial hyphae were long and fairly straight,
segmenting into chains of even, bead-like
spores. Other aerial hyphae were
tangled, branching, twisting into spirals,
also segmented into chains of bead-like
long,
spores.
Sucrose nitrate agar: Growth thin, cream-
colored, developing slowly at first, later be-
coming abundant, much folded or lichenoid;
reddish-brown to orange. Aerial mycelium
appears first over the drier edge of the
growth or in the form of thin white patches.
When the culture becomes older, a faint
bluish zone appears around the edge of the
growth. Soluble pigment faint yellowish.
Glucose-asparagine agar: Growth at first
cream-colored, becoming brownish to orange-
brown with age. Aerial mycelium white.
Soluble pigment yellowish to golden.
Yeast-glucose agar: Growth much more
rapid than in synthetic media; lichenoid,
cream to brownish. Aerial mycelium appears
at an early stage of growth, white, later
tending to become mouse-gray. Soluble pig-
ment yellowish.
Nutrient agar: Growth poor, cream-
colored to yellowish-brown to mouse-gray.
Aerial mycelium white or absent. Soluble
pigment absent or yellowish.
267
Starch Growth limited, cream-
colored, with deeper brown center. No aerial
mycelium. Limited hydrolysis.
agar:
Potato: Growth lichenoid, cream-colored
to reddish-brown. Aerial mycelium white to
gray to dark brown. Soluble pigment yel-
lowish-brown.
Gelatin: Growth cream-colored to brown-
ish. Aerial mycelium white. Slow liquefac-
tion. No soluble pigment, only a faint yel-
lowish coloration of liquefied portion.
Milk: Heavy pellicle,
colored to yellowish. Aerial mycelium gray-
ish-white. Peptonization, without coagula-
tion.
surface cream-
Cellulose: No growth.
Nitrate reduction: Positive.
Production of HS: Negative.
Antagonistic properties: Produces an anti-
bacterial antibiotic, oxytetracycline, and an
antifungal agent, rimocidin.
Habitat: Soil.
temarks: A variety of S. rimosus (forma
paromomycinus) Was briefly described (Brit.
Pat. 797,568, July 2, 1958). This variety was
isolated from a soil in South America. It dif-
fers from S. r¢mosus in certain minor cultural
properties (somewhat lighter color on agar
media) and in poorer utilization of arabinose.
Both form dense clusters of spirals on various
synthetic media and on glucose-tryptone
agar. The variety produces an antibiotic,
paromomycin, apparently closely related to
the neomycin group.
Type culture: IMRU 3558; ATCC 10,970,
209. Streptomyces rochei Berger et al.,
1949 (Berger, J., Jampolsky, L. M., and
Goldberg, M. W. Arch. Biochem. 22: 476—
478, 1949; Waksman, S. A. and Lechevalier,
H. Guide to the classification and identifica-
tion of the actinomycetes and their anti-
bioties. The Wilhams and Wilkins Co.,
Baltimore, 1953, p. 40).
Morphology: Sporophores straight, 1.5 yu
in diameter; often, but not always spirally
twisted; spirals usually short and loose with
268
rarely more than 2 to 3 coils. Spores oval to
elliptical, sometimes spherical, 1.2 to 2.8 by
OSxto. aa.
Sucrose nitrate agar: Growth thin, color-
less, covered with sandy lavender to dark
gray aerial mycelium. Reverse of growth
light gray, later becoming grayish-yellow.
No soluble pigment.
Nutrient agar: Growth cream-colored.
Aerial mycelium white. No soluble pigment.
Calcium malate glycerol: Growth good,
raised in center. Aerial mycelium gray, buff
around the edges, having a fuzzy appearance.
Glucose agar: Growth smooth, yellowish,
covered with white to gray aerial mycelium.
Yellowish soluble pigment.
Potato: Growth abundant, — lichenoid,
cream-colored. Aerial mycelium abundant,
cottony, white to gray. Color of plug be-
comes reddish-tan.
Gelatin: Cream-colored ring, covered with
white aerial mycelum. Rapid liquefaction.
Faint vellow soluble pigment.
Milk: Cream-colored to brownish ring.
Coagulation and rapid peptonization.
Starch: Growth brownish. Aerial myce-
lium mouse-gray. Diastatic action strong.
Production of HS: Negative.
Antagonistic properties: On certain com-
plex nitrogenous media, the organism shows
a wide range of antimicrobial activity,
partly because of borrelidin.
temarks: Morphologically, the culture
resembles S. albidoflavus, S. californicus, S.
lipmaniz, and certain others, but it is not
believed to be identical to any of them.
Ettlinger et al. (1958) considered this or-
ganism as a strain of S. fradiae. Okami and
Suzuki (1958) could not demonstrate any
spirals on several media tested.
Type culture: IMRU 3602; ATCC 10,739.
210. Streptomyces roseochromogenes (Jen-
sen, 1931) Waksman 1948
(Jensen, H. L. Proe. N.S.
Wales 56: 359, 1931).
Waksman and Curtis (1916) and Waks-
and Henrici,
Linnean Soc.
THE ACTINOMYCETES, Vol. II
man (1919) described an organism as A.
roseus IxXrainsky. This culture was, in con-
trast to Krainsky’s organism, chromogenic.
Jensen (1931) compared it with his own
isolates and changed the name A. roseus to
roseochromogenes, because of the fact that
the name roseus had previously been used by
Namyslowsky (1912).
Morphology: Sporophores form numerous
open and closed sinistrorse spirals; some-
times 3 to 5 branches issue together from
end point of main stem, giving impression
of brooms or verticils. Spores spherical, 1.0
to 1.2 by 1.3 to'3.0-¢ (PL. V, 2b).
Sucrose nitrate agar: Growth thin, spread-
ing, colorless to pale yellow. Aerial mycelium
pale grayish-rose.
Glucose-asparagine agar: pale
yellow. Aerial mycelium white, later be-
coming rose-cinnamon, with many small
white tufts.
Nutrient agar: Growth wrinkled, yellow-
ish-gray, later brown-red. Aerial mycelium
white, then rose-gray. Soluble pigment deep
Growth
brown.
Potato: Growth wrinkled, yellowish-gray
to grayish-black. Aerial mycelium absent or
Soluble pigment black. Melanin-
positive.
Gelatin: Colonies small, cream-colored, in
bottom of liquefied zone. Soluble pigment
brown. Liquefaction medium.
Milk: Coagulation limited; peptonization
white.
slow.
Starch media: Growth colorless, spreading.
Hydrolysis good.
Nitrate reduction: Positive.
Production of HS: Positive.
Antagonistic properties: Active against
various bacteria; produces antibiotic roseo-
mycin.
Habitat: Soil.
Remarks: Jensen (1931) obtained, on
plating the tufts of white aerial mycelium
arising on agar media, a variant with pure
white aerial mycelium.
Type culture: ATCC 13,400.
DESCRIPTION OF SPECIES OF STREPTOMYCES
211. Streptomyces roseocitreus Kato, 1953
(Kato, H. J. Antibiotics (Japan) 6A: 143;
6B: 206-208, 1953).
Morphology: Sporophores produce nu-
merous open and closed spirals of the dex-
trorse type. Spores oval, 1.2 to 1.5 by 1.6 to
1.8 p.
Sucrose nitrate agar: Growth pale olive-
buff, later changing to deep olive-buff, ivory-
yellow, or colonial buff. Aerial mycelium
scant, white. Soluble pigment at first faint
creamy, later changing to colonial buff.
Glycerol-calctum malate agar: Growth at
first transparent with gray to blackish-blue
patches, later becoming light yellowish-
olive to reed-yellow. Aerial mycelium thin,
white, at first having tinge of gray. Soluble
pigment yellowish with tinge of green.
Nutrient agar: Growth olive-buff, later
changing to deep olive-buff with bluish
patches. No aerial mycelium. Soluble pig-
ment brown.
Starch agar: Growth hyaline, cottony,
reverse becoming faint bluish. Aerial myce-
lium white, later becoming livid pink, and
finally pale grayish-vinaceous. Enzymatic
zone fair to good.
Potato: Growth thick, folded, pale olive-
buff, later deep olive to dark olive. Aerial
mycelium at first white, later becoming livid
pink to vinaceous-buff. Color of plug black-
ish-brown.
Gelatin: Whitish colonies on surface of
tube. Aerial mycelium scant, white. Soluble
pigment brown. Liquefaction slow.
Milk: Growth in yellow ring with patches.
Soluble pigment yellowish. No coagulation;
peptonization slow.
Cellulose: No growth.
Carbon utilization: Utilizes
arabinose, b-sorbitol, salicin, and sodium
sucrose, L-
acetate; not sodium succinate.
Antagonistic properties: Produces anti-
biotics roseocitrin A and B.
Habitat: Soil.
212. Streptomyces roseodiastaticus (Duché,
269
1934) nov. comb. (Duché, J. Les actinomyces
du groupe albus. P. Lechevalier, Paris,
p. 329, 1934).
Morphology: Growth consists of fine my-
celium 0.5 to 0.7 uw in diameter. Aerial my-
celium of larger diameter, but usually less
than 1 py.
Glucose nitrate agar: Growth limited,
cream-colored, becoming white with a
brownish reverse; on prolonged incubation
the culture becomes rose-gray. Soluble pig-
ment brownish.
Glycerol nitrate agar: Growth cream-
colored, becoming rose-violet; reverse red.
Asparagine agar: Growth cream-colored,
becoming rose-white; reverse of growth
reddish-brown. Soluble pigment vellow.
Tyrosine medium: Pigment
later becoming brown.
brownish,
Gelatin: Liquefied. No soluble pigment.
Melanin-negative.
Potato: Growth cream-colored, becoming
brownish-white. Soluble pigment
only in presence of glycerol.
Milk: Growth limited.
slow.
Starch: Diastatic action weak.
temarks: Closely related to S. halstedii
and considered as a transitional form. Ac-
brown,
Peptonization
cording to Ettlinger et al. (1958), this or-
ganism is related to S. griseus.
213. Streptomyces roseoflavus Arai, 1951
(Arai, T. J. Antibiotics (Japan) 4: 215-221,
1951).
Morphology: Sporophores form spirals.
Spores oval to oblong, 0.8 to 1.0 by 1.0 to
LS.
Sucrose nitrate agar: Growth colorless to
yellowish. Aerial mycelium powdery, white
to yellow-rose.
Glucose-asparagine agar: Growth color-
less to yellowish-white. Aerial mycelium
rose-colored.
Nutrient Growth folded,
white-gray to golden yellow. Aerial myce-
agar: much
Ficure 44. Verticil formation by S. roseover-
ticillatus (Reproduced from: Shinobu, R. Mem.
Osaka Univ. Ser. B, No. 5, p. 938, 1956).
lium limited to center of colonies, white to
rose.
Starch agar: Growth golden yellow. Aerial
mycelium whitish.
Potato: Growth yellow. No aerial myce-
lium. No soluble pigment.
Gelatin: Liquefaction strong. Colonies at
bottom of liquefied zone orange-brown. No
soluble pigment. Melanin-negative.
Milk: Ring cream-colored. Coagulation
and peptonization rapid, medium becoming
strongly alkaline.
Cellulose: Growth on paper fair; cellulose
decomposed.
Nitrate reduction: Positive.
Production of HS: Negative.
Antagonistic properties: Produces a basic
antibiotic, flavomycin, similar to neomycin.
temarks: Culture similar to S. micro-
flavus. Gause et al. (1957) described other
closely related cultures, such as A. roseoful-
vis.
Type culture: IMRU 3672; ATCC 13,167.
214. Streptomyces roseoverticillatus Shin-
obu, 1956 (Shinobu, R. Mem. Osaka Univ.,
B (N.S8.) 5: 84-93, 1956).
Morphology: Sporophores produce abun-
THE ACTINOMYCETES, Vol. II
dant primary and secondary verticils (Fig.
44). Spores spherical to elliptical, 0.8 to 1.0 u.
Sucrose nitrate agar: Growth thin, mod-
erate, pinkish-red. Aerial mycelium cottony,
pink to pale pink.
Glucose-asparagine agar: Growth red to
purple-red. Aerial mycelium cottony, dull
red to reddish-brown.
Nutrient agar: Growth reddish-brown to
deep brown. Aerial mycelium thin, pinkish
to red. Soluble pigment reddish-brown to
deep brown.
Potato: Growth brownish-red to dull red.
Aerial mycelium powdery, pink to reddish-
purple. Soluble pigment brown.
Milk: Growth red. Aerial mycelium pale
pink. Coagulation and peptonization strong.
Soluble pigment pale brown.
Gelatin: Rapid liquefaction.
Starch: Rapid hydrolysis.
Tyrosinase reaction: Weak.
Cellulose: No growth.
Nitrate reduction: Positive.
Carbon utilization: Utilizes fructose;
inositol uncertain. Does not utilize xylose,
rhamnose, sucrose, lactose, raffinose, or
mannitol.
Habitat: Soil in Japan.
Remarks: Resembles S. rubrireticult.
215. Streptomyces roseus (Namyslowsky,
1909; emend. Krainsky, 1914; emend. Waks-
man and Curtis, 1916) Waksman and
Henrici, 1948 (Namyslowsky, B. Centr.
Bakteriol. Parasitenk. Abt. I, Orig. 62: 564,
1909; Krainsky, A. Centr. Bakteriol. Para-
sitenk. Abt. II, 41: 682-683, 1914; Waks-
man, 8. A. and Curtis, R. E. Soil Sci. 1: 125,
1916).
Morphology: Sporophores produce num-
erous open and closed dextrorse spirals. Ac-
cording to Okami, sporophores are straight,
without spirals. Spores oval to elongate, 1.5
to 2.0 by 1.1 wu.
Sucrose nitrate agar: Growth colorless.
Aerial mycelium pale brownish-vinaceous.
No soluble pigment.
DESCRIPTION OF SPECIES OF STREPTOMYCES Prt
Glycerol malate agar: Growth colorless.
Aerial mycelium white to rose.
Nutrient agar: Growth white, turning
yellowish. No aerial mycelium. No soluble
pigment.
Starch agar: Growth colorless. Aerial my-
celium white with shade of pink. No soluble
pigment. Hydrolysis medium.
Gelatin: Growth yellowish-brown. Aerial
mycelium white. Soluble pigment brown.
Liquefaction slow; in some cases no liquefac-
tion. Melanin-negative.
Potato: Growth brownish. No aerial my-
celium. Soluble pigment brownish or absent.
Milk: No coagulation; gradual peptoniza-
tion.
Cellulose: No growth.
Invertase: None.
Nitrate reduction: Rapid.
Habitat: Soil.
Remarks: Various cultures have
described under this name. Krassilmkov
(1949) considered it as a varietal strain of
S. ruber.
Type culture: IMRU 3772.
been
216. Streptomyces ruber (KKrainsky, 1914)
Waksman and Henrici, 1948 (Ikrainsky, A.
Centr. Bakteriol. Parasitenk. Abt. II., 41:
649-688, 1914).
Not Actinomyces ruber Ruiz-Cazabo, 1894;
not Actinomyces ruber (<xruse, 1896) San-
felice, 1904.
Morphology: Sporophores straight,
branching; a few spirals may be formed.
Spores spherical and oval, 0.7 to 0.8 by 0.8
to 1.0 p.
Sucrose nitrate agar: Growth abundant,
orange to coral-red. Aerial mycelium red to
red-orange to dark red. Pigment insoluble
unless vegetable oil present in medium.
Nutrient agar: Growth elevated, wrinkled,
olive-green. No aerial mycelium.
Glucose agar: Growth abundant, coral-red.
Potato: No growth.
Gelatin: Growth yellow, flaky.
Liquefaction slow, increasing with growth.
Melanin-negative.
scant,
Milk: Dark ring with red tinge. Coagula-
tion; peptonization gradual.
Starch: Hydrolysis weak.
Sucrose: Inversion positive.
Cellulose: Growth in form of red spots.
Nitrate reduction: Positive, depending on
carbon source.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Pigments: Soluble in organic solvents;
alcohol extracts a red-violet pigment and
petroleum-ether a red-orange pigment (Kriss,
1956).
Antagonistic properties: Strongly effective
upon gram-positive bacteria. Various anti-
biotics are produced by different strains.
Habitat: Soil.
Remarks: Above description is based
largely upon that given by Krainsky. Closely
related forms include A. longisporus ruber
Krassilnikov, which is said to form some-
what longer spores, and to give sometimes a
brown coloration in protein media. A. auran-
trogriseus Gause et al. also appears to be
closely related.
217. Streptomyces rubescens (Jarach, 1931)
Umezawa et al., 1952 (Jarach. Boll. sez. ital.
soc. intern. microbiol. 3:48, 1931; Umezawa,
H., Tazaki, T., and Fukuyama, 8. J. Anti-
biotics (Japan) 5: 469, 1952).
Morphology: Sporophores short, curved,
well branched; no spirals. Spores spherical or
oval.
first
white, changing to salmon-pink. Aerial my-
Sucrose nitrate agar: Growth at
celium powdery, white. No soluble pigment.
Nutrient agar: Growth same as on sodium
nitrate agar.
Blood
growth becomes salmon-pink. Aerial myce-
agar: After 10 days’ incubation,
lium powdery, white. No soluble pigment.
No hemolysis.
Egg media: Growth colorless, changing to
coral-pink. Aerial mycelium powdery, white.
Potato: Growth coral-pink. Aerial myce-
lium powdery, white. Plug changes shghtly
272
to brown. No soluble pigment. Melanin-
negative.
Gelatin: Surface growth coral-pink. No
liquefaction. No soluble pigment.
Milk: Growth coral-pink. Aerial mycelium
powdery, white. No coagulation and no
peptonization. Soluble pigment sometimes
slightly reddish.
Starch: No hydrolysis.
Nitrate reduction: None.
Production of HS: Negative.
Carbon utilization: Glycerol and glucose
utilized, but not other carbohydrates.
Antagonistic properties: Produces an anti-
viral agent, abikoviromycin.
Remarks: A. griseoruber of Gause et al.
(1957) appears to be a closely related form.
This organism is considered by R. Gordon as
a Nocardia, related to N. asteroides.
Type culture: IMRU 3655.
218. Streptomyces rubrireticuli (Waksman,
1919) Waksman and Henrici, 1948 (Waks-
man, 8. A. Soil Sci. 8: 146, 1919).
Synonyms: Actinomyces
Waksman, 1919; A.
ed., 1925.
Morphology: Sporophores produce both
primary and secondary verticils; composi-
tion of medium influences structure of
sporophores, glucose-asparagine agar favor-
ing spiral formation. Spores oval-shaped,
smooth (PI. I a).
Sucrose nitrate agar: Growth abundant,
spreading, usually pink. Aerial mycelium
reticulus-ruber
reticulus Bergey, 2nd
white, later rose to pink.
Glucose-asparagine agar: Entire growth
abundant, spreading, rose-red.
Nutrient agar: Growth red, with yellowish
margin, becoming red. Soluble pigment dark
brown.
Starch agar: Growth white with red tinge.
Hydrolysis fair.
Potato: Growth cream-colored, later pink
to dark red. Melanin-positive.
Gelatin: Surface growth yellowish-red to
pink. Ready liquefaction. Brown pigment.
THE ACTINOMYCETES, Vol. II
Milk: Growth abundant. Coagulation and
peptonization.
Invertase: Positive.
Cellulose: Growth good.
Nitrate reduction: Rapid.
Production of H.S: Positive.
Antagonistic properties: Certain strains
produce an antibiotic designated as streptin;
others produce trichonin.
Habitat: Soil.
Remarks: Numerous cultures that pro-
duce a rose to pink substrate growth, a
soluble brown pigment in organic media, and
both primary and secondary verticils in the
sporophores have been described. It is suf-
ficient to mention A. biverticillatus by Gause
et al. (1957).
Type culture: IMRU 3631.
219. Streptomyces rutgersensis (Waksman
and Curtis, 1916) Waksman and Henrici,
1948 (Waksman, 8S. A. and Curtis, R. E.
Soil Sei. 1: 123, 1916; 8: 152, 1919).
Morphology: Sporophores produce abun-
dant close and open spirals. Spores spherical
and oval, 1.0 to 1.2 uw, with tendency to bi-
polar staming.
Sucrose nitrate agar: Growth thin, color-
less, becoming brownish to almost black.
Aerial mycelium white, becoming dull gray.
Glucose-asparagine agar: Growth abun-
dant, brown, becoming black with cream-
colored margin. No aerial mycelium appears
within 15 days.
Nutrient agar: Growth thin, wrinkled,
cream-colored.
Starch agar: Growth gray, spreading.
Hydrolysis good.
Potato: Growth abundant, much folded.
Aerial mycelium white-gray. Melanin-nega-
tive.
Gelatin: Growth cream-colored. Liquefac-
tion medium. No soluble pigment.
Milk: Cream-colored Coagulation
and slow peptonization.
Cellulose: Growth seant.
ring.
Sucrose: Inversion weak.
DESCRIPTION OF SPECIES OF STREPTOMYCES Pa les
Nitrate reduction: Good.
Production of H.S: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Various strains
produce xanthomycin-like substances; others
produce ruticin.
Remarks: The pigment formed is not
soluble. Krassilmikov (1949) considered the
organism, quite incorrectly, as a variety of
A. chromogenes.
Type culture: IMIRU 3350.
220. Streptomyces sahachirot Hata et al.,
1954 (Hata, T., Koga, F., Sano, Y., Kana-
mori, K., Matsumae, A., Sugawara, R.,
Hoshi, T., and Shima, T. J. Antibiotics
(Japan) 7A: 107-112, 1954).
Morphology: Sporophores form numerous
closed spirals with a few open spirals. Spores
oval or cylindrical, 0.7 to 1.8 by 0.5 to 0.8 uw.
Sucrose nitrate agar: Growth folded, dark
orange. Aerial mycelium velvety, white to
pale grayish-brown. Soluble pigment yel-
lowish-brown.
Calcium malate agar: Growth thin, cream-
colored with orange-yellow reverse. Aerial
mycelium thin, white, powdery. Soluble pig-
ment pale orange-yellow.
Nutrient agar: Growth glistening, white-
gray. No aerial mycelium. Soluble pigment
light brown to yellow.
Starch agar: Growth thin, yellowish-white.
Aerial mycelium thin, powdery, pale red-
brown. No soluble pigment. Slow hydrolysis.
Potato: Growth wrinkled, pale yellowish-
brown. Aerial mycelium thin, white. Soluble
pigment absent or faint brown.
Gelatin: Growth limited, white. No aerial
mycelium. No soluble pigment.
Milk: Surface growth white to pale yellow.
Coagulation; no peptonization.
alkaline. Soluble pink pigment.
Nitrate reduction: Positive.
Carbon utilization: Xylose,
lactose, trehalose, mannitol, sucrose, salicin,
glucose, maltose, mannose, glycerol, dextrin,
fructose, starch, galactose, sorbitol utilized.
Strongly
arabinose,
Rhamnose, raffinose, inositol, esculin, dulci-
tol, inulin, sodium acetate, sodium citrate.
sodium succinate not utilized.
Antagonistic properties: Produces anti-
tumor agent carzinophilin.
221. Streptomyces sampsonii (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Aerial mycelium produces
long, straight sporophores, rarely spiral-
shaped. Spores cylindrical, 0.8 to 1.0 by 0.5
m (spores oval to spherical, Waksman and
Gordon).
Sucrose nitrate agar: Growth wrinkled,
pale gray to white. Aerial mycelium very
scant, white. Soluble pigment green to buff.
Potato: Growth wrinkled, grayish. Aerial
mycelium white. Soluble pigment golden
brown (none, Waksman and Gordon).
Gelatin: Surface growth scant, gray.
Aerial mycelium trace, whitish. Liquefaction
rapid. Melanin-negative.
Milk: Surface growth good, whitish. No
aerial mycelium. No coagulation; no pep-
tonization (rapid peptonization, Waksman
and Gordon).
Starch: No hydrolysis (rapid hydrolysis,
Waksman and Gordon).
Nitrate reduction: Positive.
Tyrosinase reaction: Negative.
Temperature: 28°C,
Habitat: Potato scab.
Type culture: IMRU 3371.
222. Streptomyces sayamaensis Arishima
et al., 1955 (Arishima, M., Sekizawa, Y., Sato,
T., and Miwa, K.
29: 810-817, 1955).
Morphology: Sporophores straight, spores
short rods to cylindrical, 1.0 by 1.5 w.
J. Agr. Chem. Soc. Japan
Sucrose nitrate agar: Growth pale yellow.
Aerial mycelium gray with brownish tinge.
Soluble pigment yellow.
Glucose-asparagine agar: Growth white to
pale yellow, turning pale brown. Aerial my-
celium white, becoming pale brown.
Calcium malate Growth limited.
agar:
274
Aerial mycelium gray. Soluble pigment
brownish.
Starch agar: Growth brownish-yellow.
Aerial mycelium white, turning gray. Hy-
drolysis.
Nutrient agar: Growth pale orange-yellow.
No aerial mycelium.
Potato: Growth heavy, pinkish-gray with
purplish tinge. Soluble pigment reddish-
brown.
Gelatin: Surface pellicle pale yellow. No
soluble pigment. No liquefaction in 15 days
at 26°C. Melanin-negative.
Milk: Yellow-gray ring. Coagulation and
peptonization.
Nitrate reduction: Negative.
Cellulose: No growth.
Optimum temperature: 35-87°C.
Tyrosinase reaction: Negative.
Carbon utilization: Utilizes b-galactose,
sucrose, maltose, sodium citrate and succi-
nate; does not utilize xylose, arabinose,
lactose, rhamnose, raffinose, inulin, manni-
tol, sorbitol, inositol, and salicin.
Antagonistic properties: Produces chlor-
tetracycline.
Habitat: Soil in Japan.
temarks: Related to S. aureofaciens.
THE ACTINOMYCETES, Vol. II
999
240.
Streptomyces scabies (Thaxter, 1891)
Waksman and Henrici, 1948 (Thaxter, R.
Ann. Rept. Conn. Agr. Expt. Sta. 1891, p.
153):
Morphology: Sporophores much branched,
wavy or slightly curved; occasionally form
spirals. Spores cylindrical, 0.8 to 1.0 by 1.2
to 1.5 w (Fig. 45).
Sucrose nitrate agar: Growth abundant,
wrinkled, raised, cream-colored.
Aerial mycelium cottony, white to gray.
Growth — re-
stricted, folded, cream-colored. Aerial my-
gray to
agar:
Glucose - asparagine
celium scant, white to gray.
Nutrient agar: Growth wrinkled, white to
straw-colored, opalescent to opaque. No
aerial mycelium. Soluble pigment deep
golden brown.
Potato: Growth gray, opalescent, becom-
ing wrinkled, black. Aerial mycelium seant,
erayish-white. Color of plug brown.
Gelatin: Surface growth cream-colored,
becoming brown. Liquefaction slow. Soluble
pigment yellowish.
Milk: Surface ring brown, with greenish
tinge. Coagulation and peptonization lim-
ited.
Starch: Hydrolysis.
Figure 45. Sporophores of S. scabies, showing
of EK. Baldacci, University of Milan, Italy).
chains of transparent spores, X 15,000 (Courtesy
DESCRIPTION OF SPECIES OF STREPTOMYCES 215
Sucrose: Inversion.
Nitrate reduction: Positive.
Tyrosinase reaction: Strong.
Antagonistic properties: Certain strains
show positive antimicrobial action; others
are negative.
Habitat: Numerous strains of this organ-
ism have been isolated from various forms of
potato scab and sugar beet scab throughout
the world. True causative agent of scab.
Remarks: According to Hoffmann (1958),
growth on synthetic agar is reddish with
dark gray aerial mycelium; on glucose agar,
growth is colorless with blue-gray aerial my-
celium; on asparagine agar, growth is dark
red with no aerial mycelium.
Closely related forms include S. clavzfer,
S. spiralis, S. carnosus, and S. sampsonzi
described by Millard and Burr; also A.
xanthostromus and A. ochroleucus of Wollen-
weber. Various strains differ in the amount
of aerial mycelium produced and in their
biochemical properties.
Type culture: IMRU 3018.
224. Streptomyces setonii (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight, wavy,
formed in clumps. Spores oval, 0.6 to 0.8 by
0.85 yu.
Sucrose nitrate agar: Growth abundant,
smooth, yellow to brown. Aerial mycelium
gray to olive-buff. Soluble pigment faint
yellowish to brown.
Nutrient agar: Growth colorless. Aerial
mycelium smooth, white. Soluble pigment
brownish. Melanin-negative.
Glucose agar: Growth lichenoid, gray to
brown. Aerial mycelium abundant, white to
olive-buff. Soluble pigment golden brown.
Potato: Growth heavy, wrinkled, brown
to red-violet. Aerial mycelium abundant,
white to green to olive-buff.
Gelatin: Surface growth gray. Aerial my-
celium white. Rapid liquefaction. Soluble
pigment brownish.
Milk: Surface growth, covered with ring
of white aerial mycelium. Questionable co-
agulation, followed by rapid peptonization.
Starch cream-colored.
Aerial mycelium patchy, white. Hydrolysis.
Cellulose: Growth colorless.
Nitrate reduction: Positive.
Temperature: Grows well at 37.5°C.
Habitat: Scabby potatoes.
Remarks: Millard and Burr also described
a similar form under the name A.
agar: Growth
setonai
flavus. Ettlinger et al. (1958) consider this
organism as a strain of S. griseus. Hoffmann
(1958) described an organism with light gray
aerial mycelium as a strain of S. setoniz.
Type culture: IMRU 3375.
225. somaliensis (Brumpt,
1906; emend. Erikson, 1935) Waksman
(Brumpt, E. Arch. Parasitol. Paris 10: 489,
1906; Precis de Parasitologie. Paris, 2nd ed.,
p. 967, 1913; Erikson, D. Med.
Council (Brit.) Spee.
1935, p:. 17-18).
Morphology: Substrate growth made up of
simple branching, unicellular mycelium with
long straight filaments. Aerial mycelium
Streptomyces
Xesearch
Rept. Ser. No. 203:
forms straight nonsegmented sporophores
with typical chains of spores,
diameter.
20 M in
Glucose-asparagine agar: Growth thin,
smooth, and soft.
Glycerol nitrate agar: Growth abundant,
colorless to dark gray and black.
Nutrient agar: Growth abundant, granu-
lar, yellowish, with small discrete colonies at
margin; later growth colorless, colonies
umbilicated.
Potato: Colonies round and oval, partly
piled up in rosettes. Aerial mycelium whitish-
gray. Plug discolored. Later, aerial myce-
hum becomes transient, growth nearly black.
Blood agar: Growth in form of small, dark
found and umbilicated,
brown. colonies.
piled up in confluent bands. Reverse red-
black. Blood hemolyzed.
Dorset’s egg medium: Growth colorless,
276
becoming opaque, cream-colored, very wrin-
kled. Later, rough, yellow; medium lique-
fied.
Gelatin: Black
sediment at bottom. Rapid liquefaction.
Milk: Surface pellicle heavy, wrinkled.
Milk coagulated and completely peptonized.
Starch: Hydrolysis.
Growth cream-colored.
Habitat: Frequently found in Africa.
femarks: Although S. somaliensis has
long been known, there has been, until re-
cently, no detailed description of the or-
ganism beyond the fact that it possesses
around the grain a distinctly hard sheath
which is insoluble in potash and eau de
javelle. The rare occurrence of septa and
occasional intercalary chlamydospores 1s re-
ported by Brumpt, but has not been con-
firmed by Erikson. Chalmers and = Chris-
topherson merely mentioned the growth on
potato as yellowish-white and lichenoid,
without describing any aerial mycelium.
According to Mariat, S. somaliensis hy-
drolyzes gelatin, serum albumin, and egg
albumin; utilizes casein hydrolyzate, but not
urea, (NH,).SO, and KNO; as
sources; utilizes glucose, maltose, and fruc-
nitrogen
tose, but not xylose, starch, mannitol, or
paraffin as carbon sources.
226. Streptomyces spectabilis Dietz, 1957*
(Brit. Pat. 811,757, April 8, 1959; Am. Rev.
Tuberc. 75: 576, 1957).
Morphology: Sporophores monopodially
branched, long, straight. Pigment granules
produced in both substrate and aerial myce-
hum.
Sucrose
nitrate Growth mottled
orange to cream-orange. Aerial mycelium
agar:
mottled orange to orange.
Starch-nitrate agar: Growth cream-colored,
flecked with orange. Aerial mycelium pale
pink to orange. Starch hydrolyzed.
Starch-peptone-beef extract agar: Growth
cream-colored, turning orange. Aerial myce-
* Personal communication.
THE ACTINOMYCETES, Vol. II
lium deep orange to pale pink. Soluble pig-
ment yellow.
Gelatin: Medium liquefaction. Soluble
pigment slightly yellow to dark brown.
Melanin-negative.
Milk: Growth orange. Soluble pigment
brown. Peptonization varies with strain.
Acid formation by some strains.
Carbon utilization: Utilizes various sugars
and organic acids, depending on strain. Does
not utilize rhamnose, sucrose, inulin, ducitol,
p-sorbitol, fumarates, oxalates, or salicylates.
Nitrate reduction: Negative.
Production of H.S: Positive. Some strains
negative.
Antagonistic properties: Produces antibi-
otic streptovaricin.
Remarks: Closely related to S. fulvissemus.
227. Streptomyces spheroides Wallick et al.,
1955 (Wallick, H., Harris, D. A., Reagan,
M. A., Ruger, M., and Woodruff, H. B.
Antibiotics Ann. 1955-1956, p. 909-917).
Morphology: Sporophores form spirals,
the majority of which are closed and com-
pact; in some areas the spirals appear ball-
like. Spores oval, 0.7 to 1.1 by 1.5 to 2.0 u.
Sucrose nitrate agar: Substrate growth
white, becoming straw-colored. Aerial my-
celium abundant, white, tinged with cream
to olive-gray. No soluble pigment.
Glucose -asparagine = agar:
growth pale yellow. Aerial mycelium white,
becoming gray. No soluble pigment.
Substrate
Glucose-peptone agar: Growth moderate,
yellow. Aerial mycelium grayish-white. No
soluble pigment.
Starch agar: Growth heavy, cream- to
straw-colored. Aerial mycelium white.
Potato: Growth slow, secant, white, later
becoming heavy, gray. Aerial mycelium
gray. Soluble pigment dark brown.
Gelatin: Cream-colored, flaky sediment.
Rapid liquefaction. No soluble pigment.
Milk: Slow coagulation and peptonization.
Slight acidification,
DESCRIPTION OF
Cellulose: No growth.
Carbon utilization: No gas from adonitol,
arabinose, cellobiose, dextrin, dextrose,
galactose, lactose, levulose, maltose, manni-
tol, mannose, raffinose, rhamnose, salicin,
sucrose, or xylose.
Antagonistic properties: Produces anti-
biotie novobiocin.
Habitat: Soil.
femarks: According to Kuroya et al.
(1958), this organism is related if not identi-
cal to S. griseoflavus.
228. Streptomyces spiralis (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, S. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight or
spiral-shaped. Spores cylindrical, 1.0 to 1.7
by 0.9 uw (Fig. 46).
Sucrose nitrate agar: Growth rough or
granular, yellowish-golden. Aerial mycelium
vinaceous-buff to dark grayish-olive. Soluble
pigment pale vinaceous to fawn-colored.
Potato: Growth poor, wrinkled, grayish-
vinaceous. Aerial mycelium white to grayish-
vinaceous. Plug colored brown around and
below growth.
Gelatin: Growth limited, gray. Aerial my-
celium scant, white. Liquefaction rapid.
Melanin-negative.
Milk: Surface growth good. Aerial myce-
lium abundant, white. Coagulation and
rapid peptonization.
Starch: No hydrolysis.
Nitrate reduction: None.
Tyrosinase reaction: Negative.
Habitat: Potato seab.
Remarks: Krassilnikov (1949) considered
this organism as belonging to the A. scabies
group.
229. Streptomyces sptroverticillatus Shin-
obu, 1958 (Shinobu, R. Botan. Mag. Tokyo
71: 87-93, 1958).
Morphology: Verticil formation usually
occurs near base of aerial mycelium, but
generally not so remarkable as in the other
SPECIES OF STREPTOMYCES 2
~]
a |
FIGURE 46. Sporophores of S. spiralis, showing
that not all are transformed into spores, X 8,000
Milan,
(Courtesy of IE. Baldacci, University of
Italy).
verticil-forming species. Occasionally, very
few tufts on the skirt of the colony. Nitella
type verticils, generally primary only, seldom
About 2 to 4 radial
branches. On synthetic media, many spirals
secondary. short
in form of curled tips with | to2 turns, seldom
D
3 turns; diameter of spirals about 5 to 8 yu;
sometimes snail-like and hook-like curls.
Occasionally loose or closed spirals with 2 to
3 verticil turns, sinistrorse. Spores spheroid,
somewhat ellipsoid; about 0.8 uw in length
(Fig. 47).
Sucrose nitrate agar: Growth colorless to
Ficure 47. S. spiroverticillatus (Reproduced
from: Shinobu, R. Botan. Mag. (Tokyo) 71: 88,
1958).
pale brown to yellowish-orange. Aerial my-
celium thin, somewhat cottony, white.
Glucose-asparagine Growth pale
yellow-orange. Aerial mycelium good, cot-
agar:
tony, white to brownish.
Calcium malate agar: Growth yellow-
orange to light brown. Aerial mycelium
abundant, cottony, white.
Nutrient agar: Growth golden yellow to
buff. No aerial mycelium. Soluble pigment
pale brown.
Potato: Growth yellowish-brown to brown.
Aerial mycelium abundant, cottony, white
to brownish-white. Soluble pigment brown.
Milk: Growth yellow to yellow-orange.
Aerial mycelium poor, light brownish-gray.
Soluble pigment yellowish-orange. No co-
agulation; rapid peptonization,
Gelatin: Growth poor; liquefaction strong.
Tyrosinase reaction: Somewhat unstable,
generally positive, weak.
Diastase reaction: Fairly strong.
Nitrate reduction: Positive.
Carbon utilization: Utilizes lactose, fruc-
THE ACTINOMYCETES, Vol. II
tose, and xylose. Sucrose and inositol un-
certain. Does not utilize rhamnose, man-
nitol, and raffinose.
Habitat: Soil.
230. Streptomyces sulphureus (Rivolta, 1882
emend. Gasperini, 1894) Waksman (Rivolta,
S. Arch. path. Anat. Phys. 88: 389, 1882;
Gasperini, G. Centr. Bakt. Abt. 1, 15: 684,
1894; Waksman, 8S. A. Soil Sci. 8: 102-104,
1919).
Synonym: Actinomyces
Waksman, 1919.
Not A. sulphureus Berestnew, 1897.
This organism is usually found in culture
collections under the name of Actinomyces
bovis. Baldacci (1937, 1947) emphasized the
synonymy of this organism, listing as many
as 13 different names. The following de-
scription is based upon the data of Waks-
man (1919), who also spoke of it as A. bovis.
Sucrose nitrate agar: Growth white, turn-
ing yellow. Aerial mycelium light, powdery,
sulfur-yellow. No soluble pigment.
(Harz)
bovis
Calcium malate-glycerol agar: Growth
brownish. No aerial mycelium.
Nutrient agar: Growth at first cream-
colored, later becoming fawn-colored, brown,
then almost black. Aerial mycelium pale
yellow-green. No soluble pigment. Melanin-
negative.
Glucose agar: Growth yellowish, later be-
coming dark. Aerial mycelium thin, sulfur-
yellow.
Starch: Fair hydrolysis.
Potato: Growth abundant, much wrin-
kled, gray to canary-yellow. Aerial myce-
lium yellow, turning sulfur-yellow. Plug at
first not pigmented, later turning brownish.
Gelatin: Growth gray to brownish. No
aerial mycelium. No pigment.
Liquefaction rapid at 37°C; slow at 18°C.
Milk: Surface growth thin, yellowish.
Coagulation and peptonization.
Carbon utilization: Ready utilization of
soluble
glucose, lactose, sucrose, maltose, glycerol,
and various organic acids.
DESCRIPTION OF SPECIES OF STREPTOMYCES 279
Nitrate reduction: Positive.
Production of HS: Negative.
Invertase: None reported.
Remarks: Ettlinger et al. (1958) considered
certain strains of this organism as belonging
to the S. griseus series.
231. Streptomyces tanashiensis Hata et al.,
1952 (Hata, T., Ohki, N., and Higuchi, T.
J. Antibiotics (Japan) 5: 529-534, 1952).
Morphology: Sporophores almost straight.
Spores spherical to oval, 1.0 by 1.2 yp.
Sucrose nitrate agar: Growth grayish-
yellow. Aerial mycelium white-gray, turning
brownish-gray. Soluble pigment light yellow.
Potato: Growth brown. Aerial mycelium
dark gray to whitish-gray. Soluble pigment
dark brown.
Gelatin: Soluble pigment brown. Rapid
liquefaction.
Milk: Yellowish surface ring. Coagulation
and peptonization.
Starch: Hydrolysis. Most suitable for anti-
biotic production.
Nitrate reduction: Negative.
Tyrosinase reaction: Positive.
Production of H.S: Positive.
Optimum pH: 5.8 to 6.5.
Antagonistic properties: Produces luteo-
mycin.
Habitat: Soil.
Remarks:
antibioticus.
Resembles S. aureus and S.
232. Streptomyces tendae Ettlinger et al.,
1958 (Ettlinger, L., Corbaz, R., and Hitter,
R. Arch. Mikrobiol. 31: 351, 1958).
Morphology: Sporophores form verticils;
chains of spores as open, regular spirals.
Spores smooth (Pl. I ¢).
Glycerol nitrate agar: Growth thin, light
yellow. No aerial mycelium. No soluble pig-
ment.
Glucose-asparagine agar: Growth lhght
yellow to light carmine. Aerial mycelium
cottony, clnnamon-brown.
Calcium malate agar: Growth light yellow
to brownish-yellow. Soluble pigment brown-
ish-yellow.
Gelatin: Growth lght yellow. Aerial my-
celium sparse. Liquefaction limited. Soluble
pigment dark brown.
Starch agar: Growth thin, light yellow.
Limited hydrolysis of starch.
Potato: to dark. Aerial
mycelium powdery, chalk-white.
Milk: Growth brownish-yellow. Aerial my-
Growth brown
celium sparse. No coagulation; weak pep-
tonization.
Antagonistic properties: Produces anti-
biotic carbomycin.
Habitat: Soils in France.
Remarks: Organism said to be melanin-
negative, although dark brown pigment re-
ported on gelatin.
233. Streptomyces tenuis (Millard and
Burr, 1926) Waksman (Millard, W. A. and
Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight. Spores
cylindrical, 0.9 by 0.8 u.
Sucrose nitrate agar: Growth thin, flat,
yellowish-drab. Aerial mycelium deep olive-
buff. Soluble pigment pale orange-yellow.
Glucose agar: Growth thin, flat. Aerial
mycelium olive-buff. Soluble pigment green.
Potato: Growth good. Aerial mycelium
deep olive-buff. Soluble pigment gray to
olive to black.
Nutrient potato agar: Growth wrinkled,
grayish. Aerial mycelium white to vinaceous-
fawn. Soluble pigment golden brown.
Gelatin: Growth pale gray. Aerial myce-
lium scant, white. Rapid hquefaction. Solu-
ble pigment yellow.
Milk: Growth Aerial
white. Coagulation; incomplete peptoniza-
tion.
Starch: Hydrolysis.
Nitrate reduction: Negative.
good. mycelium
Tyrosinase reaction: Negative.
Habitat: Potato scab.
234. Streptomyces thioluteus Okami, 1952
280
(Okami, Y. Taxonomic study of antibiotic
streptomyces. Thesis, Hokkaido University,
Japan, 1952).
Morphology: Aerial hyphae with few
branches. No spirals, but verticils produced
oceasionally, depending on composition of
medium.
Glycerol nitrate agar: Growth yellowish-
brown, penetrates deep into medium. Aerial
mycelium thin, white, with dark yellowish
tinge. Soluble pigment yellowish-brown.
Nutrient agar: Growth wrinkled, yellow-
ish-brown. Aerial mycelium scant, white.
Soluble pigment slight, yellowish-brown.
Starch agar: Growth thin, cream-colored.
No aerial mycelium. No hydrolysis.
Gelatin: Growth yellowish-brown at bot-
tom of liquefied portion. No aerial mycelium.
Soluble pigment slght, yellowish-brown.
Slow liquefaction.
Potato: Growth wrinkled, cream to yel-
lowish. No aerial mycelium. Soluble pigment
slight, yellowish-brown.
Milk: Growth on surface of milk yellowish.
Aerial mycelium scant. Coagulation occurs
in 2 to 3 days, followed by slow peptoniza-
tion.
Blood agar: Growth dark gray with green-
ish tinge. Aerial mycelium dark. No hemoly-
sis.
Nitrate reduction: Negative.
Production of H.S: Negative.
Antagonistic properties: Produces anti-
fungal substance, aureothricin (Maeda,
1953).
Type culture: ATCC 12,310.
235. Streptomyces tumuli (Millard and
Beeley, 1927) Waksman (Millard, W. A.
and Beeley, fF. Ann. Appl. Biol. 14: 296-311,
1927).
Sucrose nitrate agar: Growth gray, later
becoming opaque dark. Aerial mycelium
arises at center of growth, at first white,
later becoming pale gray. Surface of growth
covered with colorless drops leaving small
black craters. No soluble pigment.
THE ACTINOMYCETES, Vol. II
Glucose-asparagine agar: Growth wrin-
kled, pale gray. Aerial mycelium white,
arising in concentric rings around a dark
bare center. Soluble pigment olive-colored.
Nutrient agar: Growth good, lustrous,
slimy, gray. No aerial mycelium. No soluble
pigment. Melanin-negative.
Potato: Growth heavy, slimy, black. No
aerial mycelium. Soluble pigment grayish-
brown.
Gelatin: Growth beaded. No aerial myce-
hum. Liquefaction rapid. No soluble pig-
ment.
Milk: Growth good; no aerial mycelium.
Coagulation and slight peptonization.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Habitat: Mound seab of mangels.
236. Streptomyces venezuelae Ehrlich et al.,
1948 (Ehrlich, J., Gottheb, D., Burkholder,
P. R., Anderson, L. E., and Pridham, T. G.
J. Bacteriol. 56: 467-477, 1948; Pridham,
T. G. and Gottheb,. D. J. Bactenol:*56:
107-114, 1948).
Morphology: Sporophores monopodially
branched, straight or shghtly and irregularly
curved. Spores oval to oblong, 0.4 to 0.9 by
0.7 to 1.6 «, smooth. (PE dlp):
Sucrose nitrate agar: Aerial mycelum
light lavender.
Nutrient agar: Substrate growth yellow
to brown. Aerial mycelium gray. Soluble
pigment dark brown to black.
Calcium malate agar: Substrate growth
vellow to brown; aerial mycelium gray.
Soluble pigment dark
Glucose agar:
brown.
Potato: Growth abundant, gray to dark
brown. Aerial mycelium gray. Soluble pig-
ment dark brown or black.
Gelatin: Liquefaction rapid. Soluble pig-
ment dark brown.
Milk: Peptonization.
dark brown.
Starch agar: Growth white to lavender.
Soluble pigment
Hydrolysis.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Nitrate reduction: Positive.
Tyrosinase reaction: Positive.
Production of HS: Positive.
Carbon utilization: Good growth: xylose,
arabinose, rhamnose, D-glucose, D-mannose,
p-fructose, bD-galactose, cellobiose, starch,
dextrin, glycerol, acetate, citrate, succinate,
and salicin. Shght or no growth: b-ribose,
sucrose, raffinose, inulin, erythritol, dulcitol,
mannitol, sorbitol, inositol, and malate. No
growth: formate, oxalate, tartrate, salicylate,
phenol, 0-cresol, m-cresol, p-cresol.
Antagonistic properties: Produces chlor-
amphenicol, an antibiotic active against
various gram-positive and gram-negative
bacteria, rickettsiae, and psittacosis group.
Habitat: Different soils.
Remarks: This organism is variable, re-
sembling in some respects S. lavendulae, al-
though Okami (1956) found it to be mark-
edly different. Krassilnikov described a form
under the name A. rectus, and a related form,
A. rectus brunneus, which belong to this
group. Gause et al. (1957) described a form
as A. venezuelae var. sprralis. S. phaeochromo-
genes var. chloromyceticus Okami is identical
with S. venezuelae.
Morais et al. (1958) described a variety of
S. venezuelae as roseosport with a_rose-
colored rather than lavender aerial myce-
lium, not producing any antibiotic and not
chomogenic on organic media.
Type species: IMRU 3534; ATCC 10,712.
237. Streptomyces verne (Waksman and
Curtis, 1916) Waksman and Henrici (Waks-
man, 8S. A. and Curtis, R. E. Soil Sci. 1: 120,
1916; 8: 156, 1919).
Morphology: According to Jensen (1931),
sporophores are long, spiral-shaped. Spores
spherical and oval.
Sucrose nitrate agar: Growth abundant,
spreading, lichenoid, glossy, yellowish, be-
coming brownish. Capacity to produce aerial
mycelium lost on cultivation.
Glucose-asparagine agar: Growth abun-
dant, lichenoid, center raised, gray with
281
purplish tinge. No aerial mycelium. Soluble
pigment faint brown.
Nutrient agar: Colonies small, grayish,
with depressed center, becoming wrinkled.
No aerial mycelium. No soluble pigment.
Potato: Growth wrinkled, cream-colored,
becoming gray. Aerial mycelium absent or
scant, white.
Gelatin: Colonies small, cream-colored.
No aerial mycelium. Soluble pigment green,
a property lost on continued cultivation.
Rapid liquefaction.
Milk: Ring pinkish-brown. Coagulation
and rapid peptonization.
Starch: Growth scant, restricted, brown-
ish; hydrolysis rapid.
Cellulose: Growth good.
Nitrate reduction: Positive.
Production of HS: Negative.
Temperature: Optimum 37°C.
Antagonistic properties: Limited activity
against some bacteria.
Remarks: Soluble green pigment produced
by freshly isolated cultures; in time, this pig-
ment becomes brown. According to Ettlin-
ger et al. (1958), this organism should be
regarded as a strain of S. olivaceus.
Type Culture: IMRU 3353.
238. Streptomyces — verticillatus — (KXriss,
1938) Waksman (Ixriss, A. Mikrobiologiya 7:
LQ5—111, 1938).
Morphology: Substrate mycelium pro-
duced by monopodial branching. Aerial my-
celium characterized by primary verticils
produced on straight The
number of verticils at the proximal ends of
the primary sterile hyphae is much larger
than in the younger portions. Secondary
verticils are also produced at the ends of the
primary. Spores cylindrical and oblong, 1.0
to 1.7 by 0.8 p.
Sucrose nitrate agar: Aerial mycelium well
developed, velvety, at first white, later dark
sporophores.
gray or gray-green.
Nutrient agar: Growth brown. No aerial
mycelium. Soluble pigment brown.
282
Potato: Soluble pigment brown.
Gelatin: Rapid liquefaction.
Milk: Coagulation and peptonization.
Starch: Hydrolysis.
Cellulose: No growth.
Nitrate reduction: Rapid.
Sucrose: Inversion.
Production of H.S: Positive.
Antagonistic properties: Weak.
Habitat: Rhizosphere of wheat grown in a
salinized soil.
Remarks: A. verticillatus viridans was de-
scribed by Krassilnikov (1941) as a substrain
of this organism.
239. Streptomyces violaceoniger (Waksman
and Curtis, 1916) Waksman and Henrici,
1948 (Waksman, 8. A. and Curtis, R. E.
Soil Sei. 1: 111, 1916).
Synonym: S. violaceus-niger.
Morphology: The sporogenous hyphae are
frequently sterile. Sporophores monopodially
branched. Waksman and Curtis (1916) re-
ported no spirals, but Ettlinger et al. (1958)
found compact spirals. Spores spherical and
oval, 1:2°to 1.5 by 1.2 to 2.34, Smooth (PI.
TL a);
Sucrose nitrate agar: Growth at first dark
eray, turning almost black. Aerial mycelium
white to gray after the colony is well de-
veloped. Soluble pigment at first bluish, later
turning almost black.
Potato: Growth at first very shght, but
after 48 hours develops into continuous,
thick yellowish-gray smear, which later
turns brown, with white aerial mycelium
covering the growth. Melanin-negative.
Gelatin: Growth gray; no aerial mycelium.
Liquefaction rapid. No change in color.
Production of H.oS: Negative.
Antagonistic properties: Produces anti-
biotic nigericin,
Habitat: Soil.
temarks: According to Ettlinger et al.
(1948) the color of the aerial mycelium is
carmine-red to cinnamon-brown; with age,
the aerial mycelium liquefies and turns black.
THE ACTINOMYCETES, Vol. II
This organism was believed to belong to the
S. hygroscopicus group. Nomi (1960) came
to similar conclusions.
240. Streptomyces violaceoruber (Waks-
man and Curtis, 1916) Waksman (Waksman,
S. A. and Curtis, R.' E. Soil Sci. 12 110=111,
1916; 8: 160-163, 1919).
This organism has an interesting history.
In the original description of Waksman and
Curtis (1916), it was listed in the text (p.
110) as A. violaceus, the word “ruber” being
left out due to poor proof-reading; in the
key, however (p. 130), as well as in the
following paper by Waksman (1919), in
which a complete description was given, it was
correctly listed as A. violaceus-ruber. The
above error was unfortunately repeated in
the first and second (p. 3874) editions of
Bergey’s Manual. In the third edition of this
manual (1930), Bergey himself changed the
name of this organism to Actinomyces Waks-
mani (p. 489). In the fourth and fifth (p.
867) editions, it was changed to Actinomyces
coelicolor (Miller) Lieske, and finally in the
sixth and seventh editions to Streptomyces
coelicolor (Miller) Waksman and Henrici.
Only the recent studies in which both or-
ganisms, S. coelicolor Miller and S. violaceo-
ruber Waksman and Curtis, were directly
compared (Kutzner, 1956; Zihner and
Ettlinger, 1957; Kutzner and Waksman,
1959) demonstrated that they are distinctly
different.
There are marked physiological and bio-
chemical differences between SS. coelicolor
and S. violaceoruber. They particularly in-
clude differences in color and morphology of
the aerial mycelium, antagonistic properties,
and pigment production. S. coelicolor is ac-
tive upon fungi and yeasts, as first shown by
Miller; several strains of S. violaceoruber
produce antibacterial antibiotics, such as
actinorhodin, coelicolorin, and mycetin. The
pigment of S. coelicolor changes to green at
an alkaline reaction, that of S. violaceoruber
to blue. The nature of the pigment has been
DESCRIPTION OF SPECIES OF STREPTOMYCES
studied by Conn (1943) and by Cochrane
and Conn (1947).
Various strains closely related to S. vio-
laceoruber have been isolated all over the
world; some have been listed as varieties,
such as achrous and flavus (Gause et al.,
1957). Most of the strains now in the culture
collections, designated as S. coelicolor, ac-
tually belong to S. violaceoruber.
Type culture: Waksman and Curtis strain
No. 3030, available in the IMRU culture
collection.
Synonyms:
A. violaceus Waksman and
(Waksman and Curtis, 1916).
A. violaceus-ruber Waksman and Curtis
(Waksman, 1918).
A. waksmani Bergey (Bergey’s Man-
ual, 3rd ed. 1930).
A. coelicolor (Miller) Lieske (Bergey’s
Manual, 4th and 5th ed., 1934, 1939).
S. coelicolor (Miller) Waksman
Henrici (Bergey’s Manual 6th
7th ed., 1948, 1957).
A. coelicolor (Miiller)
(KXrassilnikov, 1941).
A. coelicolor WKrassilnikov (Gause et al.,
1957).
Possible synonym: A.
new emend. Krassilnikoy.
Morphology: Aerial mycelium mono-
podially branched; abundant formation of
spirals with 3 to 8 turns, sinistrorse. Ac-
cording to Naganishi and Nomi (1954), two
or more sporulating branches may grow from
the same spot on the main sporophore.
Secondary branches may also be produced.
Terminal branches are often arranged in
clusters or umbellate forms. Terminal hy-
phae carry many spirals. Spores spherical to
oval, 0.7 to 1.0 by 0.8 to 1.5 » (PI. IV Ab).
Surface of spores smooth. Asporogenous,
nonpigmented strains can be obtained by
plating out cultures on carbohydrate-free
synthetic media containing nontoxic surface-
acting agents (Erikson, 1955b).
Curtis
and
and
Kkrassilnikov
pluricolor Berest-
283
Sucrose nitrate agar: Substrate growth
colorless at first, becoming red,
to dark blue. Aerial mycelium
thin, powdery, white, becoming ash-gray,
abundant,
then blue
with a bluish tinge; on some media, light
pink to cinnamon; sometimes blue drops can
be observed on the surface of the aerial my-
celium. Soluble red pigment on acid media,
changing to dark blue as medium becomes
alkaline.
Glycerol-asparagine agar: Growth good,
violet to deep blue. Soluble pigment diffuses
through medium.
Glucose-asparagine agar: Growth poor;
red pigment does not diffuse readily.
Nutrient agar: Growth white, becoming
red with white margin. No soluble brown
pigment. Melanin-negative.
Potato: Small, brownish, lichenoid colonies.
Aerial mycelium white. Mycelium and plug
gradually colored red and blue.
Gelatin: Growth cream-colored, becoming
pink or blue. Liquefaction slow.
Milk: Gray surface ring, with red or blue
tinge. Coagulation limited; peptonization
rapid.
Starch agar:
rapid.
Cellulose: Growth good.
Nitrate reduction: Excellent.
Sucrose: Inversion.
Carbon
Growth pink. Hydrolysis
sources: Utilizes L-xylose, L-
arabinose, L-rhamnose, b-fructose, raffinose
(some strains only faintly), pb-mannitol. None
or poor utilization by most strains: sucrose,
Inulin.
Antagonistic properties: Most strains do
not show any strong antagonistic effect;
several cultures, which seem to belong or are
closely related to S. violaceoruber, produce
coelicolorin, actinorhodin,
and mycetin.
streptocyanin,
Habitat: Very common, especially in field
soils.
temarks: Ettlinger ef al. (1958) considered
S. violaceoruber,
quite incorrectly, as a
234
strain of S. fradiae. Krassilnikov (1949)
considered it as a synonym of S. coelzcolor.
Type culture: IMRU 3030.
241. Streptomyces violaceus (Gasperini,
1894, emend. KKrassilnikov) Waksman (Gas-
perini, G. Centr. Bakteriol. Parasitenk.,
Abt. I 15: 684, 1894; Krassilnikov, N. A.
Actinomycetales. Izvest. Akad. Nauk. SSSR,
Moskau, p. 15, 1941).
Morphology: Aerial hyphae long, straight,
seldom
also branched
short,
branching;
THE ACTINOMYCETES, Vol. II
hyphae. Sporophores produce open, sinis-
trorse spirals with 2 to 3 coils. Spores spheri-
cal and oval (Fig. 48).
Agar media: Substrate growth lichenoid,
at first red, red-blue,
purple-violet. Some cultures produce fat
becoming finally
droplets in the colony, pigmented red or
purple. Aerial mycelium white to gray, pro-
duced poorly or not at all; some substrates,
like cellulose, paraffin, or fats, favor forma-
tion of aerial mycelium. Different pigments
Ficure 48. Sporophores and spores of S. violaceus, grown for 11 days on glucose-asparagine agar
(top, hyphae); for 8 days on yeast-glucose agar (bottom left, sporophore); for 4 days on potato agar
(bottom right, spores) X 18,500 (Reproduced from: Lechevalier, H. A. and Tikhonienko, A. 8. Mikro-
biologiya 29: 43-50, 1960).
DESCRIPTION OF SPECIES OF STREPTOMYCES
are formed in different media and under dit-
ferent conditions of growth. Pigments dis-
medium do with
solved in not
reaction.
change
Sucrose nitrate agar: Growth dark brown.
Aerial mycelium white. Soluble pigment be-
comes violet to dark violet.
Potato: Growth red-brown to
Aerial mycelium, if present, white. Soluble
pigment grayish-brown.
Gelatin: Aerial mycelium white. Liquetac-
tion slow. Soluble pigment gray-brown.
Milk: Growth-grayish brown. Coagulation
questionable; peptonization slow.
Starch: Hydrolysis weak.
Cellulose: Growth weak,
mycelium light gray.
Nitrate reduction: Positive.
Sucrose: Rapid inversion.
Pigment: According to Kriss (1936), the
pigment is soluble in water and in 96 per
cent alcohol.
Melanin: According to Hoffmann (1958),
this species is melanin-positive.
Antagonistic properties: Exerts strong
antagonistic effect upon various gram-posi-
tive bacteria.
Remarks: According to Krassilnikoyv, this
species includes A. violacea Rossi-Doria, A.
violaceus-caesart. Waksman and Curtis, Ac-
tinomyces 103 and 109 Lieske; also A.
incanescens Wollenweber and A. brasiliensis
Lindenberg (the last is probably a Nocardia).
A subspecies, A. violaceus chromogenes is also
included. Some of the cultures described by
Gause ef al. (1957) may also be included
here, such as A. lateritius, A. roseoviolaceus,
A. violaceorectus, A. viridoviolaceus, and A.
violaceus var. rubescens.
Type culture: IMRU 3497.
brown.
violet. Aerial
242. Streptomyces virgatus (Krassilnikov)
Waksman (Krassilnikov, N. A. Actino-
mycetales. Izvest. Akad. Nauk, SSSR,
Moskau, p. 32, 1941).
Morphology: Sporophores short, in form
289
of tufts. Spirals produced rarely. Spores
cylindrical, elongated; in some strains round
to oval.
Agar media: Substrate growth yellow-
green to citron-yellow or pure yellow; on
some media pale green. Pigment insoluble.
Some strains produce a brown substance in
protein media. Aerial mycelium weakly de-
veloped, white or pale yellow.
Gelatin: Liquefaction rapid.
Milk:
rapid.
Starch: Hydrolysis rapid.
Cellulose: No growth.
Coagulation and peptonization
Sucrose: Inversion.
Nitrate reduction: Positive.
Antagonistic properties: None.
Habitat: Soil.
243. Streptomyces virginiae Grundy et al.,
1952 (Grundy, W. E., Whitman, A. L.,
Rdzok, E. J., Hanes, M. E., and Sylvester,
J. C. Antibiotics & Chemotherapy 2: 399-
408, 1952).
Morphology: usually
straight; occasionally spirals are observed at
Sporophores
or near the tips of the hyphae. Spores cylin-
drical deto Wo byOs7o toweO a:
nitrate Growth
cream-colored. Aerial mycelium light gray-
ish-lavender. No soluble pigment.
Glucose-asparagine agar: Growth sparse,
cream-colored to light brown. No aerial my-
Sucrose agar: sparse,
cellum.
Calcium malate agar: Growth abundant,
cream-colored. Aerial mycelium white, be-
coming tinged with grayish-pink to lavender.
Nutrient Growth white,
turning cream-colored to light brown. Aerial
mycelium white, turning light grayish-pink
to lavender. Soluble pigment light brown.
Oatmeal agar: Growth abundant, cream-
colored, turning golden brown. Aerial my-
celium abundant, light rose, turning lavender
and gray. Soluble pigment pale yellow, turn-
agar: sparse,
ing light brown.
286
Starch agar: Growth thin, colorless. Aerial
mycelium rose to lavender-colored. Hy-
drolysis.
Potato: Growth abundant, spreading,
brownish. Aerial mycelium grayish-pink to
lavender. Browning of the potato.
Gelatin: Surface pellicle gray to brownish.
Aerial mycelium thin, white. Soluble pig-
ment brown. Liquefaction slow.
Milk: Growth brown. Coagulation none;
peptonization slow. Milk becomes dark gray-
brown or black.
Nitrate reduction: Limited or absent.
Production of H.S: Positive.
Carbon utilization: Utilizes glucose, man-
nose, galactose, maltose, starch, glycerol,
sodium acetate, sodium citrate. Does not
utilize xylose, lactose, sucrose, mannitol,
sorbitol, potassium sodium tartrate.
Antagonistic properties: Produces an
antibiotic, actithiazie acid.
Remarks: Various related organisms have
been listed. It is sufficient to mention 4A.
gobitricinit, A. roseolus, A. syringini, and A.
roseolilacinus, described by Gause et al.
(1957).
Type culture: IMRU 3651.
244. Streptomyces viridans (Krassilnikovy,
1941) (Krassilnikov, N. A. Actinomycetales.
Izvest. Akad. Nauk. SSSR, Moskau, 1941).
Morphology: Sporophores branched, spi-
ral-shaped. Spores cylindrical.
Glycerol nitrate agar: Growth olive-green
with soluble green pigment. Aerial mycelium
dark gray, olive-colored, or gray-green, vel-
vety, covering the whole growth.
Nutrient agar: Growth brown-green. Sol-
uble pigment brownish. Melanin-negative.
Potato: Growth brown. Aerial mycelum
light gray. Soluble pigment olive-green
(Hoffmann, 1958).
Gelatin: Rapid liquefaction.
Milk: No coagulation; rapid peptoniza-
tion; soluble brown pigment.
Starch: Hydrolysis rapid.
Cellulose: Growth poor.
THE ACTINOMYCETES, Vol. II
Nitrate: Reduction to nitrite.
Sucrose: Inversion rapid.
Antagonistic properties: None; some
strains are weakly active.
femarks: Related to S. intermedius.
Drechsler (1919) described two similar
strains, Nos. X and XIV. Gause et al.
(1957) described a related strain as A. roseo-
viridis.
245. Streptomyces viridis (Lombardo-Pel-
legrino, 1903) Waksman (Lombardo-Pelle-
grino, P. Riforma med. 39: 1065-1067, 1903.
Summarized by Baldacci, E. Atti ist. botan.
“Giovanni Briosi” e lab. crittogam. univ.
Pavia (Ser. IV) 11: 221-223, 1939).
Morphology: Sporophores long or short,
straight, undulated; frequently producing
broom-shaped clumps. Spores ovoid, 0.7 to
1.4 w in diameter.
Agar media: Substrate growth on all media
at first hyaline, later turning green to dark
green. Soluble pigment green. The cultures
also grow under anaerobic conditions, but
produce no soluble pigment. Aerial mycelium
on all media cottony, whitish to grayish.
Potato: Growth dark violet. Aerial my-
celium white. Melanin-negative.
Production of H.S: Positive.
Antagonistic properties: Not reported, or
negative.
Habitat: Soil.
temarks: Baldacci and Comaschi (1956)
concluded that the culture described by
Krainsky (1914) as A. griseus belongs more
accurately to the S. viridis series. According
to Hoffmann (1958), the A. griseus Krainsky
appears to belong to this group, although he
refers to it as A. griseus Krassilnikoy. It is
said to produce broom-shaped sporophores
with spirals. Growth colorless, turning light
brown. Aerial mycelium velvety, light gray
turning dark dray. No soluble pigment.
Melanin-negative. Growth on potato lichen-
oid. Milk not coagulated, but peptonized.
Gelatin liquefied. Starch hydrolyzed. It
grows on cellulose.
DESCRIPTION OF SPECIES OF STREPTOMYCES
Certain other forms belonging to this
eroup have been described, such as A. griseus
variabilis and A. griseus zonatus of Krassil-
nikov (1949), A. badius and A.
(1957).
listed viridis sterilis as a strain that lost the
malachiticus
of Gause et al. Krassilnikov also
capacity to produce aerial mycelium.
Millard and Burr (1926) described, under
A. viridis, an organism that produces dark
to black growth on sucrose-nitrate agar,
with a mouse-gray aerial mycelium, gradu-
ally black. On
growth is at first colorless, gradually becom-
becoming nutrient agar,
ing gray; aerial mycelium gray to mouse-
gray. On gelatin, it produces a thin colorless
growth and a faint brownish pigment; rapid
liquefaction.
Duché (1934) described an organism under
the name A. viridis; he later changed this
name to A. baarnensis. This organism was
isolated as a contaminant of cultures of S.
albus and S. lavendulae.
IMRU
Type culture:
Millard and Burr).
Sate (Strain: OL
237
246. Streptomyces viridochromogenes (Kra-
1948
(Xrainsky, A. Centr. Bakteriol. Parasitenk.
Abt. II., 41: 684-685, 1914).
Morphology: Sporophores monopodially
insky, 1914) Waksman and Henrici.,
branched, with numerous open or compact,
sinistrorse spirals, 3 to 5 p in diameter, oc-
curring as side branches. Spores short, oval
or spherical, 1.25 to 1.5 w (Figs. 49-51); sur-
face covered with long spines (Pl. II k).
Sucrose nitrate agar: Growth cream-col-
ored with dark center, becoming dark green;
reverse yellowish to light cadmium. Aerial
mycelium white, becoming light green to
light blue.
Glucose-asparagine agar: Growth abund-
ant, spreading, wrinkled, gray, becoming
black. Aerial mycelium appears late; white,
later becoming green to light blue.
Nutrient agar: Growth restricted, gray,
with greenish tinge. No aerial mycelium.
Soluble pigment brown.
Potato: Growth abundant, gray-brown.
Figure 49. Chains of spores of S. viridochromogenes, grown for 16 days on glucose-asparagine-CaCl»
agar, X 13,500 (Reproduced from: Lechevalier, H. .
1960).
\. and Tikhonienko, A. 8S. Mikrobiologiya 29: 43-50,
233 THE ACTINOMYCETES, Vol. II
Figure 50. S. viridochromogenes grown for 380 days on potato agar, X 13,500 (Reproduced from:
Lechevalier, H.
Aerial mycelium white.
black.
Gelatin: Surface growth cream-colored,
Soluble pigment
becoming greenish. Positive liquefaction.
Soluble pigment brown.
Milk: Surface growth dark brown; coagu-
lation and peptonization.
Starch agar: Colonies circular, spreading,
yellowish. Hydrolysis.
Cellulose: No growth.
Sucrose inversion: Positive.
Nitrate reduction: Positive.
Production of Hos: Positive.
Tyrosinase reaction: Positive.
Temperature: Optimum 37°C,
Antagonistic properties: Active upon
fungi.
Habitat: Soil.
A. and Tikhonienko, A. S. Mikrobiologiya 29: 43-50, 1960).
Remarks: This group occurs abundantly
(1958)
various forms under different names, such
in nature. Gause et al. described
as A. bicolor, A. coeruleofuscus, A. coeruleo-
rubidus, and A. coerulescens, and a variety,
longisporus. According to Ettlinger et al.
(1958), S. chartreusis also belongs to this
group.
247.
Taber,
Waksman
(Waksman, S. A.
A guide to the classification
Streptomyces viridoflavus
1953
Lechevalhier, H.
and and
and identification of the actinomycetes and
their antibiotics. The Williams & Wilkins
Co., Baltimore, 1953, p. 66).
Not A. viridiflavus Duché.
Morphology: Sporophores formed in fas-
cicles; tufts, with some curling of tips, are
produced on certain media. Tendency to lose
DESCRIPTION OF
SPECIES OF STREPTOMYCES
289
FIGuRE 51.
Milan, Italy).
property of producing aerial mycelium. Sub-
merged sporulating lateral branches form
single spores at the tips.
limited,
Aerial
Sucrose nitrate Growth
agar:
cream-colored to yellowish green.
Sporophores of S. viridochromogenes, X 30,000 (Courtesy of E.
Baldacci, University of
mycelium usually absent. No soluble pig-
ment.
Glucose nutrient agar: Growth lichenoid,
vellowish-brown to olive-brown. Aerial my-
eelium abundant, later covering the whole
290
surface of growth with a mat, yellowish to
gray. Soluble pigment brownish or absent.
Glucose-asparagine agar: Growth moist,
vellow to yellow-green. Aerial mycelium
abundant, grayish-yellow to sulfur-yellow,
later overgrown by white sporulating hy-
phae. Soluble pigment absent or faint yellow.
Nutrient agar: Growth moist, gray to
light green with green to almost bluish tinge
at bottom of slant. Nonsporulating aerial
mycelium appears much later; it is white to
gray. No soluble pigment.
Potato: Growth lchenoid, brownish to
greenish-yellow to dark olive-green. Aerial
mycelium absent, or formed as thin, yellow-
ish layer on drier portions of growth. Soluble
pigment absent or dark brown.
Gelatin: Growth in form of surface ring,
canary-yellow. Slight liquefaction. Soluble
pigment brown to dark brown, a property
that may be lost on cultivation.
Starch: Hydrolysis.
Cellulose: Limited growth, no destruction
of cellulose.
Production of H.S: Negative.
Carbon utilization: No growth with su-
crose, lactose, or rhamnose; good growth on
mannose and glucose.
Antagonistic properties: Produces an anti-
fungal substance, candidin.
Habitat: Soil.
Type culture: IMRU 3685.
248. Streptomyces viridogenes (Millard and
Burr, 1926) Waksman (S. viridis of Millard,
W. A. and Burr, 8. Ann. Appl. Biol. 13:
580, 1926).
Morphology: Sporophores long, sympodi-
ally branched, straight. Spores spherical,
0.9 u, smooth (PI. II 0).
Sucrose nitrate agar: Growth abundant.
Aerial mycelium olive-gray. Soluble pigment
greenish-yellow to blackish-green.
Glucose-asparagine agar: Growth smooth,
raised, olive-buff. Aerial mycelium abund-
ant, ight gray to deep mouse-gray. Soluble
pigment yellowish to greenish-yellow.
THE ACTINOMYCETES, Vol. II
Nutrient agar: Growth lichenoid, cream-
colored. No aerial mycelium. No soluble
pigment.
Nutrient agar with glucose: Growth gray
to black. Aerial mycelium gray. Soluble pig-
ment dark brown.
Starch agar: Growth gray to brown. Aerial
mycelium thin, white. Hydrolysis positive.
Potato: Growth gray to olive-gray. Aerial
mycelium either absent or white, turning
gray. Soluble pigment brown.
Gelatin: Growth grayish. Aerial mycelium
scant, white to gray. Liquefaction rapid.
Soluble pigment light golden brown.
Milk: Surface growth good. Aerial my-
celium scant, white. Coagulation rapid and
peptonization gradual.
Nitrate reduction: Positive.
Tyrosinase reaction: Negative.
Temperature: Grows well at 37.5°C
Habitat: Potato scab.
Yemarks: Ettlinger et al. (1958) reported
that this species is melanin-negative.
249. Streptomyces wedmorensis (Maillard
and Burr, 1926) Waksman (Millard, W. A.
and Burr, 8. Ann. Appl. Biol. 13: 580, 1926).
Morphology: Sporophores straight,
branched. Spores oblong, 0.8 to 0.9 by 0.6 to
0.8 yu.
Sucrose nitrate agar: Growth flat, thin,
grayish. Aerial mycelium white to gray.
Nutrient potato agar: Growth wrinkled,
grayish. No aerial mycelium. Melanin-
negative.
Potato: Growth wrinkled, grayish. Aerial
mycelium white. Plug pigmented drab.
Gelatin: Growth fair. No aerial mycelium.
Liquefaction medium.
Milk: Growth greenish. Coagulation and
slow peptonization.
Starch: Hydrolysis.
Nitrate reduction: Positive.
Tyrosinase reaction: Negative.
Temperature: Grows well at 37.5°
Habitat: Potato tubers.
DESCRIPTION OF SPECIES OF STREPTOMYCES
250. Streptomyces willmoret (Erikson
1935) Waksman and Henrici, 1948. (Erik-
son, D. Med. Research Council (Brit.)
Spec. Rept. Ser. No. 203: 19-20, 1935).
Morphology: Submerged growth
unicellular mycelium frequently
branched at short intervals, presenting pe-
eculiar clubbed and budding forms with
separate, round, cells.
The filaments are characteristically long,
homogeneous, and much interwoven. Aerial
mycelium profuse in most media, with a
marked tendency to produce loose spirals
with chains of ellipsoidal spores. Thick aerial
clusters may also be formed.
Glucose-asparagine agar: Growth color-
less, wrinkled, confluent, with smooth entire
large discrete colonies like flat
rosettes. Aerial mycelium scant, white.
cOon-
sists of
occasional swollen
margin;
Glycerol nitrate agar: Round, smooth,
cream-colored colonies, heavy texture, mar-
gin submerged. Stiff, sparse aerial spikes.
Peptone-beet extract or nutrient agar:
Growth heavy, colorless, lichenoid, rounded
elevation covered with white aerial myce-
lium. Later, submerged margin, round con-
fluent growth; aerial mycelium marked in
concentric zones.
Potato agar: Fair growth, partly sub-
merged. Aerial mycelium grayish-white.
Gelatin: Colonies minute, colorless. Posi-
tive liquefaction.
Milk: Coagulation and slow peptonization.
Dorset’s egg medium: Large, round, color-
less, scale-like colonies, radially wrinkled,
later growth brownish; medium discolored.
Serum agar: Smooth colorless discoid col-
onies; marked umbilication after 2 weeks.
Production of H.S: Negative.
Antagonistic properties: Positive.
Source: Streptothricosis of liver.
temarks: Ettlinger ef al. (1958) place
this species in the S. griseus series.
Type culture: IMRU 3332.
251. Streptomyces xanthophaeus Linden-
291
ybein, 1952 (Lindenbein, W. Arch. Mikro-
biol. 17: 361-383, 1952).
Morphology: No description.
Glycerol nitrate agar: Growth brownish.
Aerial mycelium white-gray or reddish-gray.
Soluble pigment yellow-brown.
Glycerol malate Growth deep
orange. Aerial mycelium white-gray to red-
gray. Soluble pigment deep orange.
Glucose-asparagine agar: Growth diffuse,
light yellow. Aerial mycelium white. Solu-
ble pigment light yellow.
Nutrient agar: Growth hight brown. Aerial
mycelium ash-gray to white. Soluble pig-
vellow-brown. Melanin-
agar:
ment yellow to
negative.
Glucose-peptone agar: Growth light yel-
low. Aerial mycelium ash-gray. Soluble pig-
ment yellow.
Starch agar: Growth Aerial
mycelium violet-gray. Hydrolysis rapid.
Potato: Growth lichenoid. Aerial myce-
lium gray. No soluble pigment.
Gelatin: Growth brown. Aerial mycelium
ash-gray. Soluble pigment yellow-brown.
Liquefaction strong.
Milk: Growth lichenoid. Aerial mycelium
gray to violet. Soluble pigment dark brown.
lichenoid.
Peptonization strong.
Cellulose: No growth.
Antagonistic properties: Produces geo-
mycin, active against gram-negative bac-
teria.
Habitat: Limestone deposit in Germany.
Remarks: Related to S. erythraeus and
S. erythrochromogenes. Kutzner (1956) stud-
ied five soil isolates. Four strains did not
form any spirals; one did. The spores were
smooth. The soluble pigment on glucose-
peptone agar was dark brown. He thus con-
sidered this species as melanin-positive.
Addendum
After the text of this volume was com-
pleted, the following newly described forms
appeared in print:
Streptomyces aerocolonigenes Shinobu and
292
Kawato (Botan. Mag. Tokyo 73: 212-216,
1960).
Actinomyces aureoverticillatus KrassiInikov
and Dzi-Shen (Mikrobiologiya 29: 482-489,
1960).
Streptomyces herbaricolor Kawato and
Shinobu (Mem. Osaka Univ. B 8: 114-119,
1959).
THE ACTINOMYCETES, Vol. II
Streptomyces massasporeus Shinobu and
Kawato (Botan. Mag. Tokyo 72: 853-854,
1959).
Streptomyces — ostreogriseus (Antibiotic
E-129) Brit. Pat. 799,053, July 30, 1958.
Streptomyces psammoticus Virgilio and
Hengeller (Farmaco, Ed. Sci. 15: 164-174,
1960).
Chapter 9
The Genus Micromonospora
The genus J/icromonospora is character-
ized by the production in nutrient media of a
well developed substrate mycelium, 0.2 to
0.6 w in diameter, partly penetrating into
the medium. The substrate or vegetative
hyphae are straight or curved, branching,
without cross walls. Aerial mycelium is not
formed at all or only in a rudimentary, non-
sporulating form, when the hyphae arise
upward directly from the substrate myce-
hum.
Multiplication occurs by means of frag-
ments of mycelium and special spores formed
singly. A swelling takes place at the end of
the sporophore; later the swelling is sepa-
rated by a cross wall, giving rise to spherical,
oval, or oblong spores, 1.0 to 1.5 by 0.8 to
1.2 uw. The sporophores are often branched,
each branch forming a spore at the end,
giving rise to a grape-like bunch of spores.
These germinate in a manner similar to the
spores of Streptomyces. The mycelium and
spores are gram-positive, not acid-fast (Fig.
52).
The colonies of J/icromonospora are simi-
lar to those of Streptomyces. They are com-
pact, leathery, smooth or lichenoid, raised or
flat. They are frequently colored red or
orange or yellow, occasionally brown or green
to almost black or blue. The pigments, ex-
cept the dark brown, are not dissolved into
the medium.
In characterizing species of J/icromono-
spora, T’ao Ho and Potter (1960) considered
morphological properties as primary criteria
for identification of the organisms. The
293
important physiological characteristics in-
cluded the disintegration of cellulose, in-
version of sucrose, and the reduction of
nitrate. The investigators emphasized that
the color of the growth and the form of the
colony could not serve as basic characteris-
Certain may different
colors for the mass of growth and for the
tics. strains show
spores. Reproducibility of colony color for a
given organism could not be obtained on the
same medium. The color itself was not con-
sistent, varying through every shade of yel-
low, orange, pink, red, and brown. Many
species gave more than one colonial form.
The large spores of JZ. globosa were very
helpful in differentiating it from J/. fusca or
AM. chalcea.
Micromonospora species are aerobic or
anaerobic, and mesophilic. They grow
readily at 25-40°C. Thermal death point of
the mycelium is 70°C in 2 to 5 minutes;
spores resist 80°C for 1 to 5 minutes. They
utilize various carbon and nitrogen sources,
both organic and inorganic (Fig. 53).
The type species is icromonospora chalcea
(Foulerton) Orskoy.
The genus MJicromonospora comprises nine
species, which can be classified as follows:
Classification of the genus Micromonospora
A. Aerobic.
I. Sporophores long.
1. Sporophores showing little branching.
a. No aerial mycelium.
2. Micromonospora chalcea
b. Rudimentary aerial mycelium.
6. Micromonospora gallica
294
2. Sporophores form extensive branching.
a. Growth colorless; brown spores
appear in mass.
7. Micromonospora globosa
b. Growth pigmented.
al. Growth green to dark green.
a2. Spores blue.
3. Micromonospora coerulea
b?. Spores black or brown.
1. Micromonospora bicolor
b!. Growth pink to orange-colored.
a2. Pigment not excreted into
substrate.
8. Micromonospora parva
b?. Red-brown pigment excreted
into substrate.
5. Micromonospora fusca
IL. Sporophores short.
1. Growth brown; spores dark brown.
4. Micromonospora elongata
B. Anaerobic.
9. Micromonospora pro-
pronica
Various other micromonosporas have been
observed in natural substrates, but either
have not been isolated or only insufficiently
Ld ‘
*
FIGURE 52. Micromonospora (clumps of spores)
growing in a compost.
THE ACTINOMYCETES, Vol. II
te
af
re ~ F
e \
53. Micromonospora (double spores)
FIGURE
growing in a compost.
studied. This is true, for example, of the
cellulose-decomposing, facultative anaerobic
form studied by Meyer, 1934 (Prevot, 1955);
and of J. cabaelli Maquer and Comby
(Prevot, 1955). It is also true of some of the
forms reported by Waksman et al. (1939).
Some of the micromonosporas (J/. mono-
spora and AM. vulgaris) have been placed,
because of their temperature optima, among
the thermophilic forms.
Description of Micromonospora Species
1. Micromonospora bicolor WKrassilnikov,
1941 (KKrassilInikov, N. A. Actinomycetales.
Izvest. Akad. Nauk. SSSR, 1941, p. 131).
Morphology: Sporophores long, branch-
ing, 10 to 25 uw; spores oval, 1.0 to 1.2 by
0.8 wu.
Synthetic agar: Growth green, smooth;
covered with a dark brown to black hue of
spore-bearing hyphae. Pigment insoluble.
Nutrient agar: No growth.
Potato: No growth.
Gelatin: No growth. No liquefaction.
Milk: Unchanged.
Starch: Not liquefied.
THE GENUS MICROMONOSPORA 295
Sucrose: Inverted.
Cellulose: Good growth and decomposi-
tion.
Carbon sources: Glucose, sucrose, levulose,
acetic and citric acids.
Nitrogen sources: Ammonium salts and
nitrates.
Habitat: Soil.
2. Micromonospora chalcea (Foulerton,
1905) Orskov, 1923 (Foulerton, A. Lancet
1: 1200, 1905; Orskov, J. Investigations into
the morphology of the ray fungi. Levin and
Munksgaard, Copenhagen, 1923).
Morphology: Grows well on all media,
especially = on
Growth heavy, compact, raised, pale pink to
deep orange, not spreading much into the
medium. Hyphae thin, branching,
nonseptate. Surface of growth smooth or
folded, dull or shining. Spore layer well de-
veloped, moist and glistening, brownish-
black to greenish-black; color sometimes
spreading through the whole mass of growth.
Spores oval or spherical, formed individually
on relatively nonbranching sporophores (Fig.
54).
Gelatin: Liquefaction positive. No soluble
elucose-asparagine — agar.
long,
pigment.
Milk: Coagulation and peptonization
positive.
Starch: Hydrolyzed.
Cellulose: Rapid decomposition.
Chitin: Decomposed.
Nitrate reduction: Positive.
Sucrose: Inverted.
Proteolytic action: Strong.
Temperature: Optimum for growth, 30—
35°C. Thermal death point of mycelium,
70°C in 2 to 5 minutes. Spores resist 80°C
for 1 to 5 minutes.
Source: Soil, lake mud, and other sub-
strates.
3. Micromonospora coerulea Jensen, 1932
(Jensen, H. Proce. Linnean Soc. N.S. Wales
Bio. 1932).
Morphology: Growth smooth, lustrous,
4
a
« >
L c S
~~
y vag *
jp ,
ba’ 5. re be
} ys :
<e ’
Se * i ’ 4
~»6 4 “
FIGuRE 54.
growing in a compost.
Micromonospora (single spores)
greenish-blue; pigmentation only on free
admission of Pigment insoluble.
Surface of colonies hard and glossy; thin,
oxygen.
white veil on surface resembles aerial my-
celium. Spherical blue spores produced
on branching short sporophores.
Nutrient media: Slow growth.
Liquid media: Growth at bottom, in the
form of firm, round, white to pink granules.
Gelatin hquefaction: Rapid.
Milk: Positive coagulation, but very shght
peptonization.
Starch: Hydrolyzed.
Cellulose: Not decomposed.
Nitrate reduction: None.
Sucrose: Not inverted.
Source: Occurs rarely in soil.
4. Micromonospora elongata Wrassilnikoy,
1941 (Krassilnikov, N. A. Actinomycetales.
Izvest. Acad. Nauk. SSSR, 1941, p. 130).
Morphology: Sporophores short (2 to 3
u), little branched. Spores oval, 1.0 to 1.3 by
0.8 uw (Fig. 55).
Agar media: Growth poor, adhering to
substrate in form of minute pale yellow
smooth colonies. Surface is dark brown.
Potato, gelatin, and milk: No growth.
296
Ficure 55. Micromonospora spores)
growing in a compost.
(single
Cellulose: Good growth and decomposi-
tion.
Sucrose: Inverted.
Nitrate reduction: Positive.
Habitat: Soil.
5. Micromonospora fusca Jensen, 1932
(Jensen, H., Proc. Linnean Soc. N.S. Wales,
ad: 178, 1932):
Morphology: Growth heavy, compact,
orange-colored, rapidly changing to deep
brown and nearly black. Spore layer moist,
glistening, grayish to brownish-black. Spores
oval or spherical.
Gelatin: Liquefaction slow. Soluble pig-
ment very slight.
Milk: No coagulation, slow peptonization;
grayish-brown discoloration of milk.
Starch: Hydrolyzed.
Cellulose: Limited decomposition.
Nitrate reduction: Positive or negative.
Sucrose: Inverted.
Antagonistic properties: Produces the
antibiotic micromonosporin.
Source: Soil.
6. AMicromonospora gallica (Krikson, 1935)
Waksman (Erikson, D. Med. Res. Council,
London, Pub. No. 4582, 1935, p. 24).
Morphology: Aerial hyphae retarded and
rudimentary. Typical single spores produced.
Sucrose nitrate agar: No growth.
Glycerol agar: No growth.
THE ACTINOMYCETES, Vol. II
Glucose agar: No growth.
Potato agar: Growth pale pink, moist,
granular.
Potato: Growth slow in form of pink,
translucent tending to
umbilicated and heaped up.
Ege media: Colonies minute, becoming
confluent, tangerine-colored.
Blood agar: Colonies minute, discrete,
pink. No hemolysis.
Gelatin: Growth scant, irregular, pink.
Liquefaction slow.
Broth: Pinkish flakes. Small rounded red
granules at bottom.
Milk: Surface ring yellowish-pink. Positive
colonies, become
coagulation and peptonization.
Habitat: Isolated from blood culture.
7. AMicromonospora globosa WKrassilnikovy,
1939 (Ixrassilnikov, N. A. Mikrobiologiya 8:
179, 1939; Actinomycetes. Izvest. Akad.
Nauk, SSSR, Moskau, 1941, p. 129).
Morphology: Spores spherical, 1.0 uw, ar-
ranged in clusters on long branching sporo-
phores.
Agar media: Growth at first colorless,
leathery, lichenoid; covered with a dark
brownish tarnish of spores.
Gelatin: Liquefaction slight.
Milk: Coagulated and peptonized.
Starch: Hydrolyzed.
Cellulose: Satisfactory growth.
Nitrate reduction: Positive.
Sucrose: Inverted.
Source: Soil.
8. Alicromonospora parva Jensen, 19382
(Jensen, H. Proc. Linnean Soc. N.S. Wales
af: 177, 1932).
Morphology: Growth pale pink to orange,
compact. Substrate mycelium thin, spread-
ing widely into the agar. Sporulation scant,
giving rise to thin, grayish, moist crusts on
the surface. Spores oval; in mass gray-
colored.
Gelatin: Liquefied slowly.
Milk: Unchanged; or coagulated and
slowly peptonized with faintly acid reaction.
Starch: Hydrolyzed.
THE GENUS MICROMONOSPORA
Cellulose: No decomposition.
Nitrate reduction: None.
Sucrose: Not inverted.
Source: Soil.
9. Micromonospora propionica Hungate,
1946 (Hungate, R. E. J. Bacteriol. 51: 51-
56, 1946).
Morphology: Grows very slowly. White
spherical colonies produced on
Colony consists of gradually expanding hol-
low shell, its outer surface consisting of sub-
strate hyphae, the adjacent inner portion
containing numerous spores, and the center
relatively devoid of protoplasm.
cellulose.
297
Media: Highly complex media required.
Cellulose: Good growth. Good decomposi-
tion.
Oxygen demand: Obligate anaerobe.
Carbon sources: Glucose and cellulose are
utilized.
Fermentation products: Carbon dioxide,
acetic and propionic acids.
Temperature: 30-40°C.,
Habitat: Gut of termites, rumen of cattle.
Prévot (1957)
species as an anaerobic Actinomyces, which
Remarks: considers. this
he includes in the genus Actinobacterium.
t=)
Chapter 10
The Genus Waksmania (Microbispora)
The genus Waksmania (Microbispora) 1s
characterized by the paired spores produced
on aerial hyphae (Fig. 56).
The fine mycelium (about 1 uw in diameter)
is differentiated into (a) primary or substrate
mycelium which grows into, and forms a
compact layer on top of agar media, and (b)
secondary or aerial mycelium which arises
from the primary mycelium but grows into
the air, away from the agar surface. The
substrate mycelium does not bear spores of
any type; the aerial mycelium bears spores
which are formed in longitudinal pairs. The
spores are produced either directly on the
aerial hyphae or on sporophores which
branch from the aerial hyphae.
The sporophores may be so short that the
spores appear to be produced directly on the
mycelium. The aerial mycelium forms a bud
at the side, and later the bud, or occasionally
the tip of the side branch, swells and is
separated by a cross wall giving rise to two
spherical or oval conidia, 1.4 to 1.6 w in
diameter (Fig. 57).
The germination of the spores and the
structure of substrate mycelium are similar
to those of Streptomyces.
The type species is Waksmania rosea
Lechevalier and Lechevalier.
Waksmania rosea Lechevalier and Leche-
valier, 1957 (Lechevalier, M. P. and Leche-
valier, H. J. Gen. Microbiol. 17: 104-111,
1957).
Synonym: Microbispora rosea Nonomura
and Ohara, 1957 (Nonomura, H. and Ohara,
Y., J. Ferm. Technol., 35: 307-311, 1957).
Morphology: The dominant form of this
organism on some media, after 14 days at
30°C, may consist of chlamydospores.
Hyphae do not segment, even in old cultures.
“Fairy rings,” or alternating areas of aerial
mycelium with zones which have none, occur
on some media. ‘“‘Coremia-like”’ aggregations
of aerial hyphae are formed on a variety of
substrates. Small branches are produced
monopodially in respect to the main axis of
the aerial sporogenous hyphae. Spores are
borne at the tip of these branches or sporo-
phores. Spores are spherical, 1.5 to 2.0 yu,
usually about 1.7 to 1.8 uw. Spores are borne
terminally on sporophores, as well as at the
tips of the main sporogenous hyphae and
branches. They are also borne directly on
the sporogenous hyphae. Spores are formed
in pairs and, when mature, are very easily
detached from the sporophore and from each
other.
Agar media: Growth nutrient
agar. Color of substrate growth pale pink to
coral-orange, changing to chestnut-brown.
Diffusible pigment very slight. Aerial myce-
lium white, powdery, with tendency to form
fairy rings on some media.
Sucrose nitrate agar: Growth thin, yel-
lowish-white. Aerial mycelium scant. Some
malformed spores appear.
Glucose-asparagine agar: Growth meager,
colorless. No aerial mycelium.
Yeast-glucose agar: Growth
glistening, becoming dark brown and con-
voluted. No aerial mycelium.
Oatmeal agar: Orange-pink vegetative
growth. Aerial mycelium white, with hght
pink spores. Earthy odor.
slow on
white-tan,
298
THE GENUS WAKSMANIA (MICROBISPORA)
Potato: Growth dark reddish-brown.
Aerial mycelium has a slight trace of white.
Gelatin: Growth on the bottom of tube
white, fluffy. Liquefaction slight.
Milk: Growth on the bottom of the tube
white. Surface colonies orange-pink, at-
tached to sides of test tube. Coagulation
none; peptonization complete after 1 month.
No change in pH.
Starch: Not hydrolyzed.
Sucrose: Not inverted.
Cellulose: Attacked to a very limited de-
eree.
Nitrate reduction: Negative.
Temperature: The organism is a meso-
phile, growing well at 25—-35°C. It produces
only sparse growth at 40°C, and does not
grow at all at 55°C.
Antagonistic properties: None.
Habitat: Soil. A culture of this organism
causing pericarditis and pleuritis has been
Ficure 56. Waksmania rosea, schematic repre-
sentation of the formation of aerial mycelium and
spores (Reproduced from: Lechevalier, M. P. and
Lechevalier, H. J. Gen. Microbiol. 17: 108, 1957).
299
Friaure 57. Waksmania (Microbispora) rosea: A.
sporulation; B. chlamydospores; C. germination
of conidia (Reproduced from: Nonomura, H. and
Ohara, Y. J. Fermentation Technol. 35:307, 1957).
recently isolated by Louria and Gordon
(1960).
temarks: Thiamine and biotin are es-
sential for growth; biotin also controls pig-
mentation. Ammonium compounds, nitrates,
and urea are not utilized as
nitrogen. Asparagine, glutamic acid, and
peptone are good sources.
Type cultures: IMRU 3748, 3757.
Nonomura and Ohara (1960a) found the
genus (Microbispora) widely
distributed in soils of Japan. When a par-
ticle of soil was placed on soil-extract agar in
a Petri dish, colonies of this group of organ-
isms appeared around the soil particle after
a few weeks incubation at 30°C. Five species
and two varieties were recognized, including
M. amethystogenes, M. amethystogenes var.
nonreducans, M. parva, M. chromogenes, M.
sources of
Waksmania
diastatica, and AM. rosea var. nonnitritogenes.
A system of classification of these species
and varieties was proposed, based upon their
erowth on different media, nitrate reduction
to nitrite, production of soluble pigment, and
hydrolysis of starch. Some of the strains pro-
duced violet crystals, insoluble in water but
soluble in benzene. Thiamine was required
for growth of all strains, biotin to a limited
extent.
Chapter
11
Thermophilic Actinomycetes
Our knowledge of the thermophilic actino-
mycetes dates back to the early beginnings
of general microbiology. The first students
of the microbiological population of soils and
composts observed that some of the organ-
isms found among the bacteria, actinomy-
cetes, and fungi were able to grow at much
higher temperatures than the great majority
of the members of these groups.
Globig (1888) was the first to isolate from
the soil thermophilic actinomycetes, capable
of growing at 52-65°C. Rabinowitsch (1895)
and Tsiklinsky (1903) isolated similar cul-
tures from manure, and Noack (1912) iso-
lated them from hay. Numerous other 1so-
lations of thermophilic actinomycetes were
made, from ordinary soil by Gilbert (1904),
from desert sand, feces, air, and peat, as
well as from human
sewage. It has also been observed repeatedly
that composted manure, when it has attained
intestines and from
a high temperature, or hay which has been
allowed to heat in composts, becomes cov-
ered with small white patches of fungus-like
growth. Miehe (1907) remarked that the
appearance of these patches is similar to a
coat of lime and is due to actinomycetes.
Similar observations of thermophilic com-
posts have been made by various other in-
vestigators.
Tsiklinsky inoculated potato slices with
soil or with manure and incubated them at
Do made agar
plates after 16 hours. Two cultures were
53-
Isolations were on
thus obtained. One produced chains of spores
and was considered to be, therefore, a true
actinomycete of the type now designated as
300
Streptomyces. The other formed round or
ovoid spores at the end of side branches, by
the swelling of the tips. This organism was
believed to be widely distributed in nature
and was named Thermoactinomyces vulgaris.
Because of its manner of spore formation,
this form was believed to belong to the group
of actinomycetes designated by Mrskov
(1923) as AMicromonospora, and was, there-
fore, classified by Waksman as J/icromono-
spora vulgaris. It grew at 48-68°C, with an
optimum at 57°C, and no growth at 70°C. It
remained inert for a month at 37°C or at
lower temperatures, but it became active
within 24 hours at 56-57°C. The spores were
said not to be destroyed at 100°C, even after
20 minutes. The organism grew readily on
most ordinary media; it was proteolytic but
not amylolytic. The Streptomyces, on the
other hand, was weakly proteolytic, and the
spores were less resistant to heat (Fig. 58).
Gilbert cultivated from various
several strains of a thermophilic actino-
thermo-
soils
mycete, which he designated as A.
philus. Growth on potato was much folded,
white, later becoming gray on the surface;
the plug was darkened by some cultures.
The optimum temperature was 55°C; no
growth took place at 60°C. Most strains
ceased to grow at 45°C, whereas some could
be adapted to grow on agar media at 37°C
and even at 22°C. The colonies on agar were,
after 24 to 48 hours, small, folded, lght
vellow with a dark-colored center. Gelatin
was only slowly liquefied.
Miehe considered hot composts and not
the natural actino-
‘
(a
soils as substrates of
THERMOPHILIC
mycetes. The spores of these organisms were
found to lose their vitality rapidly, especially
but they survived on hay
on agar media,
particles. One culture was designated as A.
thermophilus Berestnew; it grew best at
40-50°C, more slowly at 30°C, and not at
all at 25°C and at 60°C. The manner of
spore formation of this organism suggested
its resemblance to J/icromonospora. Miehe
also reported, however, that he saw thermo-
philic actinomycetes arith formed spores
according to the manner described by Gil-
bert. This suggests the probability that he
had representatives of the two different
groups. Schiitze (1908) found, in decompos-
ing clover hay, representatives of these two
thermophilic actinomycetes, which
appeared to belong to the Streptomyces
group and the other to the J/zcromonospora.
Several methods for isolation and cultiva-
tion of thermophilic actinomycetes have
been described by Henssen (1957). Uridil
and Tetrault (1959) suggested the incorpora-
tion of colloidal silica in a highly proteinace-
one ot
ous medium for the growth of these organ-
isms.
The various thermophilic forms have often
been classified under the common name of
‘Actinomyces thermophilus.’? Waksman et al.
(1939) demonstrated, by direct microscopic
studies, that these organisms are very abun-
dant in high temperature composts of stable
manures and plant residues. Six distinct
types or species were recognized, belonging
to two genera, one now known
myces and the other as
Two of the first group (S. thermophilus and
as Strepto-
AMicromonospora.
S. thermofuscus) and three of the second
(M/. vulgaris, M. chalcea, and M. were
isolated and cultivated. Henssen (1957) con-
fuse a)
firmed these observations and created sey-
eral new genera and species, to include these
and other forms (Fig. 59).
Kosmatchev (1959) emphasized the need
for separating the thermophilic from the
mesophilic actinomycetes, since the former
at and the latter cannot be
TOW GAC.
ACTINOMYCETES
301
FicureE 58. First photograph of a Thermoactino
myces (Reproduced from: Tsiklinsky, P. Ann.
inst. Pasteur 13: 500-505, 1899).
adapted to grow at that temperature or
under thermophilic conditions. The thermo-
philic actinomycetes form a sharply defined
ecological and widely distributed group of
microorganisms, and retain. their
philic under laboratory condi-
tions. They were believed to comprise in-
thermo-
properties
dependent species. He suggested, however,
that no specific thermophilic genera
recognized, but that they should be included
among the mesophilic forms.
be
Henssen (1957) proposed a distinct system
for classifying thermophilic actinomycetes.
This system is used, with certain modifica-
tions, in this treatise.
Classification of Thermophilic Actino-
mycetes
A. Substrate mycelium unseptated.
duced on
Spores pro-
aerial mycelium only.
they
completely transformed into spore chains
Streptomyces Waksman and Henrici
Aerial hyphae branched; are almost
302
THE ACTINOMYCETES, Vol. II
Fraure 59. Four types of thermophilic actinomycetes (Reproduced from: Henssen, A. Arch. Mikro-
biol. 26: 379, 1957).
Il. Long sterile aerial hyphae forming single
spores or chains of spores on side branches.
1. Spores produced singly on simple or
branched sporophores.
Thermomonospora Henssen
2. Spores produced in two’s or in longer
chains.
Thermopolyspora Henssen
III. Single spores or chains of spores originate
directly from substrate mycelium, which
sometimes emerges from the agar surface
like an arch.
Thermoactinomyces Tsiklinsky
B. Substrate mycelium septated. Spores formed
from both aerial and substrate mycelium.
Pseudonocardia Henssen
Genus Streptomyces
XVI. Series Thermophilus
Spores produced in chains, comparable
to the mesophilic species of Streptomyces.
I. Sporophores and chains of spores straight.
a. Aerial mycelium white to light gray.
2. Streptomyces rectus
b. Aerial mycelium white; thermotolerant.
1. Streptomyces casei
Il. Sporophores and chains of spores not straight.
1. Spore chains spiral-shaped.
6. Streptomyces thermovulgaris
2. Spore chains bent or curved.
a. Aerial mycelium white to violet-gray.
5. Streptomyces thermoviolaceus
b. Aerial mycelium gray to lavender.
4. Streptomyces thermofuscus
ce. Aerial mycelium white to light gray.
3. Streptomyces thermodiastaticus
1. Streptomyces casei (Bernstein and Mor-
ton, 1934) nov. comb. (Bernstein, A. and
Morton, H. E. J. Bacteriol. 27: 625, 1934).
Morphology: Sporophores straight, 0.5 to
0.7 w in diameter.
Agar media: Growth colorless to white.
Aerial mycelium white.
Gelatin: Liquefaction rapid.
Milk: Positive coagulation and peptoniza-
tion.
Starch: No hydrolysis.
Nitrate reduction: None.
Temperature: Optimum 40-60°C. Highly
resistant to higher temperatures and to dis-
infectants. Thermal death point 100°C.
Habitat: Pasteurized cheese.
Xtemarks: IKrassilnikov (1949) placed this
THERMOPHILIC ACTINOMYCETES
culture in the same group with A. znvul-
nerabilis (Acosta and Rossi)
1897, the latter said to be even more re-
sistant to high temperatures and to disin-
Berestnew,
fectants.
2. Streptomyces rectus Henssen, 1957
(Henssen, A. Arch. Mikrobiol. 26: 373-414,
1957).
Not A. rectus Krassilnikov.
Synonym: Streptomyces thermophilus.
Morphology: Sporophores straight, 36° to
60 uw long. Spores round or oval, 0.9 to 1.2 wp.
Sucrose nitrate agar: Growth moderate.
No aerial mycelium.
Glycerol-asparagine agar: Growth good.
Aerial mycelium moderate, white to light
eray.
Meat-—extract agar: Growth good. Aerial
mycelium light gray. Soluble pigment brown.
Cellulose-dextrin agar: Growth more or
less heavy. Aerial mycelium white-gray.
Potato: Growth colorless. Soluble pigment
brown.
Starch: Slow hydrolysis.
Nitrate reduction: Positive.
Milk: Coagulated; not peptonized in 9
days.
Gelatin: Not lquefied (Waksman et al.
(1939) obtained liquefaction).
Habitat: Fresh horse manure.
Remarks: Thermotolerant mesophile.
Grows equally well under aerobic and an-
aerobic conditions.
The name ‘“‘thermophilus” for this species
was not recognized by Henssen (1957), since
it was first used by Berestnew (1897) for
another actinomycete, apparently also a
species of Streptomyces, which was distin-
guished from S. rectus by spiral-forming
chains of aerial spores, gray or dark green
aerial mycelium, and yellow to dark brown
Noack (1912)
ganism, under the name ‘“‘thermophilus,”’
which produced a soluble red pigment. Miehe
(1907) and Schtitze (1908) also described
organisms under this name.
colonies. deseribed an_ or-
305
3. Streptomyces thermodiastaticus (Bergey,
1919) nov. comb. (Bergey, D. H. J. Bac-
teriol. 4: 301, 1919).
Morphology: Sporophores form spirals.
Spores spherical to oval, 0.9 by 0.7 or 0.8 wu.
Synthetic agar: Growth colorless. Aerial
mycelium well developed, white.
Potato: Growth brownish. Aerial myce-
hum light gray.
Gelatin: Liquefaction slow.
Milk: No coagulation; no peptonization.
Starch: Strong hydrolysis.
Cellulose: Growth good.
Nitrate reduction: Positive.
Sucrose: Inverted.
‘Temperature: Optimum, 65°C.
Habitat: Mouth of rabbit. Soil.
4+. Streptomyces thermofuscus (Waksman
et al., 1939) nov. comb. (Waksman, S. A.,
Umbreit, W. W., and Cordon, T. C. Soil Sci.
47: 49, 1939).
Morphology: Aerial mycelium gives rise
to spiral-shaped sporophores; spores spher-
ical.
Sucrose nitrate agar: At 28°C, growth
poor, deep gray, with but little aerial my-
celium. At 50°C, growth black to violet,
with gray to lavender aerial mycelium. Sol-
uble pigment brown.
Potato: Growth abundant,
ored. Aerial mycelium, none or a few white
patches. Soluble pigment black.
Gelatin: Liquefied. At 50°C, a grayish
ring is produced and a soluble pigment is
formed. At 28°C, there is growth without
any soluble pigment.
Milk: Peptonized.
Starch: Hydrolysis.
Temperature: Good growth at 50 and
60°C. Will grow at 65°C. Faint growth at
28°C.
Habitat: Horse manure.
brown-col-
Xtemarks: This species is characterized by
brown-colored aerial mycelium on synthetic
media, spiral-shaped sporophores, and abil-
ity to grow readily at 65°C.
5304
5. Streptomyces thermoviolaceus Henssen,
1957 (Henssen, A. Arch. Mikrobiol. 26: 373-
414, 1957).
Morphology: Substrate mycelium shghtly
branched. Sporophores curved, 20 to 40 u
long. Spores oval, 1.0 to 1.2 by 1.2 to 1.6 yu.
Facultative aerobe.
Synthetic agar: Growth slight. Sporo-
phores ochre-brown. Aerial mycelhum white
to violet-gray.
Glycerol-asparagine agar: Growth yellow.
Aerial mycelium abundant. Soluble pigment
violet.
Nutrient agar: Growth good. No aerial
mycehum.
Cellulose-dextrin agar:
Aerial mycelium produced.
Potato: Growth good, black-violet. Aerial
white. Soluble pigment dark
Growth — shght.
mycelium
violet.
Starch: Rapid hydrolysis.
Nitrate reduction: Negative.
Milk: Coagulation rapid; peptonization
rapid.
Gelatin: Growth orange-yellow. Lique-
faction positive. Soluble pigment produced.
Habitat: Fresh horse or swine manure.
temarks: This species was divided by
Henssen into two subspecies: (a) pingens,
producing a violet pigment on potato; (b)
apingens, producing no pigment on potato.
6. Streptomyces thermovulgaris Henssen,
1957 (Henssen, A. Arch. Mikrobiol. 26: 373—
414, 1957).
Morphology: Mycelium shows _ little
branching. Sporophores produce spirals, 20
to 40 w long. Spores oval, 1.0 to 1.2 by 1.1 to
1.5 uw. Facultative aerobe.
nitrate agar:
Aerial mycelium present or absent.
Sucrose Growth — shght.
Glycerol-asparagine agar: Growth mod-
erate to good. Aerial mycelium white to
black-violet.
Nutrient agar: Growth good, colorless.
Aerial mycelium violet-gray. No
pigment.
soluble
THE ACTINOMYCETES, Vol. II
Cellulose-dextrin agar: Growth
Aerial mycelium produced.
Potato: Growth good in places. Aerial
mycelium white to gray in spots. Soluble
pigment often black.
Starch: Rapid hydrolysis.
Nitrate reduction: Positive.
Milk: Rapidly coagulated and peptonized.
Gelatin: Liquefaction varies.
Temperature: Grows well at 40—-50°C,
somewhat better under anaerobic than
aerobic conditions. At 28°C growth slight;
at 60°C growth moderate. No aerial myce-
lium at 28°C; shght aerial mycelium at 60°C.
Habitat: Fresh compost and fresh horse
manure.
good.
Genus Thermomonospora
The type species is Thermomonospora fusca
(Waksman et al.) Henssen.
These produce
yellow growth on agar media. Substrate
mycelium nonseptated. The aerial myce-
lium is sharply lmited and white. Aerial
hyphae are simple or branched, developing
organisms colorless or
as side or terminal branches on the substrate
hyphae. The spores are formed singly on
simple or branched sporophores on the un-
branched aerial hyphae. Spore formation is
acropetal. Gram-positive. Not acid-fast.
Thermophilic, facultative aerobes.
I. Spore-masses produced in form of a head or
a bunch of grapes.
2. Thermomonospora fusca
II. Spore-masses spiked or entangled.
3. Thermomonospora lineata
III. Spores mostly single on simple or branched
sporophores.
1. Thermomonospora curvata
1. Thermomonospora curvata Henssen,
1957 (Henssen, A. Arch. Mikrebiol. 26: 373-
414, 1957).
Morphology: Aerial hyphae 30 to 50 u
long. Mass of spores clumped or hairy.
Spores round, 1.2 to 1.8 u.
Agar media: Colonies colorless or yellow.
THERMOPHILIC ACTINOMYCETES
Aerial mycelium chalk-white. Cultures are
thermophilic, facultatively aerobic. Aerial
spores produced in 3 to 4 days in hanging
drops. Spores formed on simple or branched
sporophores. Spores oval, later round.
nitrate Growth colorless.
Aerial mycelium moderate.
Glycerol-asparagine agar: Growth abun-
dant, yellow. Aerial mycelium produced.
Nutrient Growth yellow.
Aerial mycelium thick.
Potato: Growth yellow, partly covered
with aerial mycelium. Soluble pigment light
Sucrose agar:
agar: good,
brown.
Starch: No hydrolysis.
Nitrate reduction: Weak.
Milk: Unchanged in 16 days.
Gelatin: Not liquefied.
Temperature: Optimum growth at 50°C.
Limited growth at 28 and 65°C.
Habitat: Fresh cow manure and manure
compost.
2. Thermomonospora fusca (Waksman et
al., 1939) Henssen (Henssen, A. Arch.
Mikrobiol. 26: 373-414, 1957).
Morphology: Aerial hyphae 20 to 30 u
long. Spores round 1.5 to 2.0 uw. Since the
spores are not produced on the substrate
mycelium as in J/icromonospora, but ex-
clusively on the aerial mycelium, the species
was transferred from J/icromonospora fusca
to Thermomonospora fusca. The branching
of the substrate mycelium is monopodial.
The hyphae are long, straight, and form
straight side branches. The branching is so
characteristic, as compared to all the other
thermophilic species, that this species can
sasily be recognized. The aerial mycelium
is colored brown (Fig. 60).
Sucrose nitrate agar: Growth at 28°C
deep gray; at 50°C, growth is dark brown to
violet. Aerial mycelium gray to lavender.
Soluble pigment brown.
Gelatin: Liquefied.
Potato: Growth brown-colored. No aerial
mycelium. Soluble pigment black.
305
FIGuRE 60.
duced from:
401, 1957).
Thermomonospora fusca (Repro-
Henssen, A. Arch. Mikrobiol. 26:
Milk: No coagulation; slight peptoniza-
tion.
Habitat: Horse manure.
Starch: Hydrolysis.
Nitrate reduction: Slight.
Cellulose: Growth good.
Temperature: Growth and aerial myce-
lium formation are good at 50-65°C.,
9
3. Thermomonospora lineata Henssen,
1957 (Henssen, A. Arch. Mikrobiol. 26: 373-
414, 1957).
Morphology: Sporophores straight, 50 to
80 » long; spores round, 1.5 to 2.0 uw. Spore
chains hairy or clumpy (Fig. 61).
Culture was isolated on nutrient agar but
was not obtained in pure state. Aerial hy-
phae mostly branched. Spores produced on
simple or branched sporophores. Substrate
hyphae monopodial and branched. Hyphae
are not as long as in the case of 7. fusca.
Side branches are more compact.
Optimum growth: 50—-60°C.
Habitat: Composted sheep manure.
Genus Thermopolyspora
Growth on agar media colorless to yellow.
Substrate mycelium not septated. Aerial
mycelium limited, white. Aerial hyphae not
branched, developing in the form of side or
terminal branches of the substrate hyphae.
Spores produced in short chains on un-
Spore formation
branched — sporophores.
acropetal. Spore chains unbranched, straight,
306 THE ACTINOMYCETES, Vol. II
‘te
ESS
REA
Ve
»
| os
Ficure 61. Thermomonospora lineata (Repro-
duced from: Henssen, A. Arch. Mikrobiol, 26:
401, 1957).
bent or spiral-shaped. Gram-positive, non-
acid-fast, thermophilic, facultatively aerobic.
This genus comprises two species: 7.
with made up of double
bispora chains
spores, and 7. polyspora with more than
two spores.
Usually only the side branches of the
substrate mycehum grow into aerial hyphae.
The chains of spores are produced from the
unbranched sporophores around the aerial
hyphae.
Type species:
Thermopolyspora bispora
Henssen.
1. Thermopolyspora bispora Henssen, 1957
(Henssen, A. Arch. Mikrobiol. 26: 373-414,
1957).
Morphology: Aerial hyphae 20 to 30 u
long, with 20 to 40 spore chains. Spores are
double, round, 0.9 to 1.3 yp.
Cultural characters: Colonies mostly col-
orless. Aerial mycelium chalk-white. Organ-
ism thermophilic, facultatively aerobic.
The following properties are discussed on
the basis of anaerobic growth at 60°C.
Sucrose nitrate agar: Sparse growth.
Glycerol-asparagine agar: Growth shght
or abundant. No aerial mycelium.
Nutrient agar:
cehum thick.
Starch agar: No hydrolysis.
Good growth. Aerial my-
Gelatin: Unchanged.
Potato: Individual colonies without aerial
mycehum.,
Nitrate reduction: Negative.
Milk: Not changed after 7 days.
Habitat: Fresh cow and swine manure or
composted sheep manure.
Type culture: IMRU 3759.
2. Thermopolyspora polyspora Henssen,
1957 (Henssen, A. Arch. Mikrobiol. 26: 373-—
414, 1957).
Morphology: Aerial hyphae 45 to 50 u
long, 1.0 to 1.8 uw thick. Sporophores straight
to spiral-forming, individual or in groups.
Spores round, 3 to 10 by 1.1 to 1.8 wu.
Physiological properties of spore-free cul-
60°C
vellow,
conditions:
dirty
tures, at under aerobic
Colonies aerial mycelium
white.
Glycerol-asparagine agar: Growth orange-
vellow.
Cellulose-dextrin agar: Growth colorless.
Nutrient agar: Growth yellow.
Potato: Growth yellow.
temarks: These cultures resemble,
through their colored compact colonies, the
spore-free cultures of /zeromonospora. Spiral
formation has resulted in the confusion of
T. polyspora with species of Streptomyces.
Genus Thermoactinomyces
The genus Thermoactinomyces is similar im
some respects to the genus J/7cromonospora;
THERMOPHILIC ACTINOMYCETES 307
it is distinct from it, however, in the forma-
tion of a typical aerial mycelium.
Henssen emended this genus as follows:
Colonies on agar
orange. Substrate mycelium not septated.
Aerial mycelium not sharply delimited;
white or bluish-green. Aerial hyphae are
simple or branched, formed as terminal or
side branches; they may also be curved;
they grow upwards from the substrate my-
celium. Spores, single or in chains, remain
on the unbranched aerial hyphae. The or-
ganisms are thermophilic, capable of grow-
ing at 50-65°C. They are aerobic or faculta-
tively aerobic. Some have their optimum at
60°C.
Type species: Thermoactinomyces thalpo-
philus Waksman and Corke.
The genus can be classified as follows:
colorless or yellow to
I. Aerial mycelium white.
a. No soluble pigment.
6. Thermoactinomyces vulgaris
b. Soluble wine-colored to rose pigment in
certain media.
3. Thermoactinomyces thalpophilus
II. Aerial mycelium grayish-green.
2. Thermoactinomyces monosporus
III. Aerial mycelium secant, white to bluish-green.
a. No soluble pigment.
1. Thermoactinomyces glaucus
b. Soluble pigment green.
5. Thermoactinomyces viridis
IV. Aerial mycelium white to dark gray.
4. Thermoactinomyces thermophilus
1. Thermoactinomyces glaucus Henssen,
1957 (Henssen, A. Arch. Mikrobiol. 26:
373-414, 1957).
Morphology: Spore chains simple or
branched, about 7 w long, containing 4 to
10 spores. The chains are straight or bent.
Spores round or oval, 0.9 to 1.4 by 0.5 to
eo) in.
Cultural properties: Growth almost color-
less; aerial mycelium white to bluish-green.
Facultative aerobe.
Sucrose nitrate agar: Growth moderate.
Aerial mycelium moderate.
Glycerol-asparagine agar: Growth moder-
ate. Aerial mycelium scant.
Nutrient agar: Growth sparse.
Cellulose-dextrin agar: Growth good.
Aerial mycelium abundant, white to green.
Cellulose decomposed very actively.
Gelatin: Unchanged.
Potato: Individual colonies without aerial
mycelium.
Starch: Slow hydrolysis.
Nitrate reduction: Positive.
Milk: Slowly coagulated and peptonized.
Habitat: Composted sheep manure.
2. Thermoactinomyces monosporus (Leh-
mann and Schtitze) Waksman (Schiitze, H.
Arch. Hyg. 67: 35, 1908; after Krassilnikov,
1941).
Morphology: Substrate hyphae about 1.0
uw in diameter. Oval spores 1.5 to 1.8 by
1.0 to 1.4 « produced singly.
Agar media: Growth yellowish, compact,
smooth or lichenoid. Aerial mycelium gray-
ish-green. Good sporulation of hay infusion-
peptone agar; somewhat less on glycerol-
peptone and lactose-peptone agar;
peptone-glucose agar.
Potato: No growth.
Gelatin: Liquefaction positive.
none on
Milk: No coagulation or peptonization.
Blood serum: Good, smooth growth; lique-
faction positive.
Temperature: Optimum 37-55°C; grows
poorly at 27°C and not at all at 60°C.
Habitat: Self-heated hay.
temarks: Henssen (1957) considers this
species as more closely related to the genus
Thermomonospora.
3. Thermoactinomyces thalpophilus Waks-
man and Corke, 1953 (Waksman, 8S. A. and
Corke, C. J. Bacteriol. 66: 377, 1953).
Morphology: Spores, produced singly or
in short chains, are round, 0.8 to 1.2 yu.
Agar media: Grows equally well on a
variety of media under aerobic and anaero-
bic conditions. Colonies colorless to orange.
308
Aerial mycelium white. Soluble wine-red
pigment produced on sugar-containing salt
media.
Sucrose nitrate agar: Very little growth.
Glycerol-asparagine agar: Growth abun-
dant. Aerial mycelium formed, orange.
Nutrient agar: Growth yellow.
mycelium limited.
Gelatin: Some liquefaction.
Potato: Growth limited. Aerial mycelium
limited.
Starch: Rapidly hydrolyzed.
Nitrate reduction: Positive.
Milk: Coagulation rapid; peptonization
rapid.
Temperature: Grows well at 50—-60°C,
lesser growth at 40°C; no growth at 28 and
65°C.
Habitat: Manure compost.
Aerial
4. Thermoactinomyces thermophilus (Be-
restnew, 1897) nov. comb.
Morphology: Sporophores straight (No-
ack, Waksman ef al.) or spiral-shaped
(KXrassilnikov). Spores spherical.
Cultural properties: Substrate growth
yellow-brown. Soluble pigment brown. Aerial
mycelium white to dark gray.
Svnthetic agar: Growth colorless. Aerial
mycelium thin white. No soluble pigment.
Potato: Growth yellowish. No aerial my-
celium. Soluble pigment brown.
Gelatin: Liquefaction rapid. No pigmen-
tation.
Milk: Coagulation and peptonization.
Starch agar: Growth yellowish. Aerial
mycelium powdery, Starch
rapidly hydrolyzed.
Nitrate reduction: Rapid.
Cellulose: Shght growth.
Temperature: 35-55°C. Optimum 50°C.
temarks: Noack deseribed, under this
name, organisms of a bright red color, with
a red soluble pigment. Optimum 40-59°C,
Miehe and Schiitze also described organisms
white-gray.
under this name.
THE ACTINOMYCETES, Vol. II
et al., 1956 (Schuurmans, D. M., Olson, B.
H., and San Clemente, C. L. Appl. Micro-
biol. 4: 61-66, 1956).
Morphology: Spores borne singly; oval
(about 1.0 by 1.8 uw). Diameter of hyphae
approximately 0.5 pu.
Glucose-asparagine agar. No growth.
Calcium malate agar: Colonies colorless,
1 to 2 mm in diameter; a few colonies with
blue-green aerial mycelium.
Nutrient agar: Growth wrinkled, colorless,
and close to agar surface. Aerial mycelium
blue-green. Soluble pigment emerald-green.
Glucose-peptone agar: Colonies colorless,
1 to 2 mm in diameter. No aerial mycelium.
Nutrient broth: Floeculent cream-colored
submerged growth; surface growth blue-
green. Soluble pigment green.
Gelatin: Liquefied.
Potato: No growth.
Milk: Coagulation positive, followed by
peptonization.
Nitrate reduction: Negative.
Starch: Hydrolysis.
Temperature: Minimum, 37°C; optimum,
55°C; maximum, 60°C.
Antagonistic properties: Produces the
antibiotic thermoviridin, active primarily
against gram-positive bacteria.
Habitat: Composted manure pile.
temarks: Description of growth and bio-
chemical reactions after 14 days’ incubation
FIGURE 62. Pseudonocardia thermophila (Repro-
duced from: Henssen, A. Arch. Mikrobiol. 26:
5. Thermoactinomyces viridis Schuurmans 409 , 1957).
THERMOPHILIC
at 45°C. The organism in many ways re-
sembles Thermoactinomyces monosporus. The
points of difference, however, are considered
significant, the latter possessing hyphae
with a diameter of about 1 uw, an optimum
growth range of 37 to 55°C, and failing to
coagulate milk.
6. Thermoactinomyces vulgaris Tsiklinsky,
1899 (Tsiklinsky, P. Ann. inst. Pasteur 13:
500, 1899).
Synonym: J/icromonospora vulgaris Waks-
man et al., 1939; Erikson, 1953.
Morphology: Substrate mycelium fine, 0.5
u in diameter. Spherical and oval spores are
borne singly at the ends of short branches,
from which they are easily broken. They
often appear to sit directly on mycelium.
Sucrose nitrate agar: Growth colorless.
Aerial mycelium white.
Nutrient agar:
celium white.
Potato: Growth good.
Gelatin: Liquefaction positive.
Milk: Coagulation and peptonization.
Starch: Hydrolysis positive.
Cellulose: No decomposition.
Nitrate reduction: Negative.
Sucrose: Not inverted.
Temperature: Grows at 48-68°C; opti-
mum at 57°C,
Source: Human and animal excreta, high
temperature composts, self-heated hay, soil.
Remarks: Resembles mesophilic members
of the genus JJ/icromonospora, except that it
produces an aerial mycelium which forms
single spherical spores. The aerial phase of
the development of this organism is believed
to be intimately associated with its thermo-
philic nature (Erikson, 1952, 1953, 1955a).
Oxygen concentration has an important
effect upon the growth of the aerial mycelium
of this organism (Webley, 1954). The effect
of composition of medium and the growth-
temperature relationships of this organism
were studied recently by Tendler (1959).
Growth good. Aerial my-
ACTINOMYCETES
309
Genus Pseudonocardia
Substrate mycelium septated. Spores pro-
duced in substrate and in aerial mycelium.
On the basis of its morphology, Pseudo-
nocardia should be placed between Strepto-
myces and Norcardia. Along with Nocardia,
it has the common property of septation of
the substrate mycelium, but no fragmenta-
tion. In common with Streptomyces, it pro-
duces aerial mycelium which is thicker than
the substrate mycelium, and which changes
into long spore chains. It differs from Strep-
tomyces in being unable to hydrolyze gelatin
or starch.
Type species: Pseudonocardia thermophila
Henssen.
Pseudonocardia thermophila Henssen, 1957
(Henssen, A. Arch. Mikrobiol. 26: 373-414,
1957).
Morphology: Substrate sep-
tated. Spore formation in substrate myce-
mycelium
hum. Aerial hyphae unbranched, formed as
side branches of the substrate hyphae.
Spores basipetal. Substrate spores 2.5 by
1.5 to 1.8 uw. Aerial spores produced in chains
at the tip of the hyphae are 2.5 uw long; spores
produced at the base of the chain are 5 u
long by 1.5 to 1.8 mw wide. Thermophilic,
facultative aerobe (Fig. 62).
Sucrose nitrate agar: Growth yellow.
Aerial mycelium limited, white.
Glycerol-asparagine agar: Growth moder-
ate, yellow. Aerial mycelium produced,
white.
Nutrient agar: Growth good, yellow.
Aerial mycelium in thick colonies.
Potato: Colonies individual, yellow. No
aerial mycelium. Soluble pigment yellow.
Gelatin: Not liquefied.
Starch: No hydrolysis.
Nitrate reduction: Positive.
Milk: Unchanged in 16 days.
Habitat: Fresh horse manure.
Ch aww: te
12
Actinoplanaceae
Substrate mycelium usually inconspicu-
ous, formed in water, on a variety of plant
and animal materials. Aerial mycelium
usually lacking; only certain species pro-
duce such mycelium, thus resembling
Streptomyces. Reproduction by spores formed
in sporangia. The spores in Actinoplanes
possess flagella and are motile. The spores
of Streptosporangium are without flagella
and are nonmotile. Many species produce
aerial spores. These organisms can be culti-
vated on a variety of artificial media; they
will then, resemble in their growth other
actinomycetes. The family is widely dis-
tributed in soil and in fresh water. The
Actinoplanaceae can be classified as follows:
I. Aerial mycelium usually not formed;
coiled conidiophores lacking; sporangio-
spores motile.
Genus I. Actinoplanes
II. Aerial mycelium abundant; coiled co-
midiophores as well as sporangia
formed in some species; sporangiospores
are
nonmotile.
Genus II. Streptosporangium
Genus Actinoplanes Couch
(Couch, J. N. J. Elisha Mitchell Sci. Soe.
66, 87, 1950; 71, 48, 1955. Trans. N. Y.
Acad. Sei. 16, 315, 1954).
Occur on sterilized leaves in water, form-
ing a@ very inconspicuous mycelium which
branches throughout the leaf tissue. The
external hyphae are scattered or in tufts on
310
the leaf surface and form a fringe around
the edge of the leaf. Aerial mycelium is
lacking or sparingly formed; usually pinkish
to reddish, sometimes hyaline; frequently
decolorizes the green leaf and gives it a
pinkish or reddish color. Hyphae slightly to
considerably branched, irregularly coiled,
twisted or straight, sparingly septate, 0.2 to
2.6 w in diameter. Sporangia usually abun-
dant on leaves, formed only when the leaf is
at or close to the surface of the water, 2.e.
formed typically only in air, and appearing
black under the low power of the microscope,
owing to refraction; of varied sizes and
shapes. Spores in coils, nearly straight
chains, or irregularly arranged, in sporangia;
1 to 1.5 win diameter, globose or subglobose,
usually shghtly angular, with one to several
shiny bodies, with several polar flagella, and
motile; germination by a minute germ tube
which branches to form a mycelium. Spo-
rangial wall evanescent or persistent (Fig. 63).
The organisms form on various nutrient
agars a brilliantly colored, tough to pasty
growth. Surface very variable: smooth and
even with the agar or elevated bumpy,
convoluted, ridged, folded, cracked, ete.,
usually moist and shiny, rarely pulverulent.
Hyphae of two more or less distinct forms,
the submerged and the surface hyphae, the
latter usually more or less vertical and in
some species forming a compact ‘‘palisade.”’
Sporangia abundant on some agars, usually
formed at the surface. Spores formed in
some species. On certain agars, the mycelium
of some species breaks up, when crushed,
ACTINOPLANACEAE oll
into irregular pieces of hyphae, rods and
coccoid bodies.
Aerobic, gram-positive, and acid-fast.
Occur saprophytically in soils and in fresh
water, and are world-wide distribution.
Over 120 cultures were isolated.
The Actinoplanes is readily
tinguished from
leaves, the latter produces a conspicuous
aerial mycelium which that in
most species of Streptomyces, whereas no
such mycelium is usually found Actino-
The isolates of the latter grow much
genus dis-
Streptosporangium. On
resembles
planes.
more vigorously on agar than do those of
Streptosporangium. The striking dif-
ference is that in Actiznoplanes the sporangio-
spores are motile, whereas in Streptosporan-
gium they are nonmotile.
Under certain conditions of culture, some
species of Actinoplanes resemble J/icro-
monospora. A nonsporangial strain of Ac-
tinoplanes might easily be confused with
certain micromonosporas. The
AMicromonospora, however, are tormed singly
or in grape-like clusters but never in chains,
whereas in Actinoplanes they are formed
most
spores of
Ficure 63.
Lechevalier,
Actinoplanes
Institute of Microbiology).
(Prepared by H.
singly and also in chains but not in grape-
like clusters. In most species of /icromono-
certain
surface turns black, wher
not occur in
spora, on agars, the sporulating
sas this change does
Actinoplanes. In general, the
species of /icromonospora are less vigorous
in growth than those of Actinoplanes.
Several species of Actinoplanes, when
grown on potato-glucose and certain other
agars, Will form a small pasty culture which,
when mounted and crushed under
slip,
a cover-
breaks up into minute spheres, irregular
rods, and short, branched, hyphal segments,
much as in Nocardia. Such growth, however,
is not the normal condition for any species
of Actinoplanes. None of the 25 species
of Nocardia examined by Couch formed
sporangia when grown either on any of the
agars most favorable for sporangial forma-
tion or on Paspalum leaves in water.
Gaertner (1955) isolated cultures of Ac-
tinoplanes trom soil and found them capable
of decomposing keratin.
The type species
pinensis Couch.
Actinoplanes philip-
Actinoplanes philippinensis Couch, 1950
(Couch, J. N. J. Elisha Mitchell Sci. Soe.
66, 87, 1950).
Morphology: Produces a very delicate,
hyaline to pinkish-buff internal mycelium
and an inconspicuous external fringe of
threads around the entire edge of the leaf of
sterile Paspalum grass in water. Sometimes a
compact mound or tufts of hyphae are
scattered over the top surface, giving the
leaf a speckled or finely powdered appear-
ance. Hyphae are 0.5 to 1.5 uw thick,
branched, sparingly septate. Sporangia,
usually formed abundantly on grass after
about 10 days, on long unbranched sins,
mostly spherical when mature, 8.4 to 22 wu.
Spores arranged, at maturity, in
irregularly in the sporangium, about 1 to
They are discharged through a pore
or by the partial dissolution of the sporangial
wall, and swim vigorously.
coils
a12 THE ACTINOMYCETES, Vol. II
nitrate agar: Growth at room
temperature poor to fair, flat or shghtly
elevated. Margin smooth or scalloped, sec-
toring frequent. Color pale buff to tawny,
changing in some old cultures to brown with
Sucrose
a lighter margin. Forms a compact surface
layer, made up mostly of distinct palisades,
and a submerged region of loosely arranged
hyphae. Surface region frequently stratose in
old cultures, with narrow, orange-colored
layers. Sporangia fairly abundant in some
cultures, not formed in others; spherical to
irregular; frequently beneath the surface in
old cultures, owing to overgrowth by pali-
sade hyphae. Sometimes a new layer of
sporangia forms over the first layer. Odor
shghtly fragrant. Usually colors the agar
pale yellow.
Glucose-asparagine agar: Growth good to
very good, consisting of a central area of
elevated, fine convolutions, radial ridges or
bumps, and a smooth area with radial
grooves gradually sloping into the submerged
margin. Surface moist-appearing and glossy.
Color of center apricot-orange to brown,
surrounded by an ochraceous-salmon— or
FiGuRE 64. Streptosporangiwm
soc. Tis 152, 1955):
roseum (Reproduced from: Couch, J. N.
J. Elisha Mitchell Sei.
ACTINOPLANACEAE 3
light ochraceous-salmon margin. Sporangia
usually on the smooth areas, none on the
elevated parts; formed on palisade hyphae.
Potato-glucose agar: Growth good to
very good. Central area with coarse con-
bumps and_ irregular
ridges separated by radial grooves which
slope to the smooth distinct margin. Surface
volutions or large
glossy. Color apricot-orange to russet, be-
coming gray in old cultures. Soluble pigment
darkens the agar. Sporangia formed on the
margin of some cultures, absent in most.
Palisades formed.
Nutrient agar: Growth fair. Center
slightly elevated and with a wide flat margin.
Color ochraceous-orange to clnnamon-rutous.
Sporangia rarely formed.
hyphae usually not distinct.
Gelatin: Liquefied.
Habitat: Sou from Philippine Islands;
also found in African soils and in marshland
soils in Germany.
Remarks: This species is characterized
by the predominantly spherical sporangia
usually on unbranched stalks, the
rather poor and usually flat growth on
synthetic agar, and the very distinct palisade
hyphae on this medium. The dark brown
diffusible pigment on potato-glucose agar 1s
also characteristic.
very Palisade
long
Genus Streptosporangium Couch
Occurs on sterilized leaves of Paspalum
grass in water, forming an inconspicuous
mycelium which overgrows the leaves, and
an aerial mycelium which grows in scattered
or concentrically arranged tufts. The aerial
mycelium is white to pinkish on the leaves;
hyphae are much branched, sparingly sep-
tate, and about 0.5 to 1.2 uw in diameter. On
some media, sporangia are formed abun-
dantly in the aerial mycelium. Spores are
abundant in the sporangia, without flagella,
and are nonmotile. Growth poor to good on
a variety of semisolid media (Fig. 64).
Four cultures, representing three distinct
WW
—
“
species, were found to comprise this genus.
Two of the species were isolated from soil
by the soil dilution method and the third
from dog manure.
The
roseum Couch.
type species is Streptosporangium
Streptosporangium roseum Couch, 1955
(Couch, J. N. J. Elisha Mitchell Sei. Soc. 71
148, 1955).
Morphology: Grows on_ sterile leaves,
either in soil water or on damp sterile soil,
forming a substrate mycelium which spreads
over the surface of the leaf, not penetrating
or decolorizing it; it also spreads over the
soil. Aerial mycelium white at first, changing
to pale pink; it appears as single hyphae or
as minute tufts which grow to form mounds
up to 2 mm across, arranged more or less in
concentric circles. Sporangia first appear on
scattered single hyphae, apical on the main
thread or on short, lateral branches, a few
to many sporangia on one hypha. The
sporangia are white in small groups, pink in
large masses; spherical, 7 to 19 uw in diameter.
Shortly after their formation, spores are
visible as a single coil in each sporangium;
when completely formed, they are irreg-
ularly arranged. Immersion of the mature
FIGURE 65.
Streptosporangium isolated from
forest litter (Reproduced from: Van Brummelen,
J.and Went, J. F. Labor. Microb. Univ. Amster-
dam 23: 391, 1957).
314 THE ACTINOMYCETES, Vol. II
sporangium in water brings about the swell-
ing of an intersporal substance, causing
the wall and the spores to push out on one
side, forming a cone-shaped — projection
about half as long as the diameter of the
sporangium. The spores are forcibly ejected
through an opening in the cone. They are
nonmotile, spherical, 1.8 to 2.0 uw in diameter,
with one shiny globule. Sporangial wall is
persistent for several hours after spore
discharge. In addition to sporangia, spores
are also formed in coils somewhat as in
Streptomyces, though the coils are much less
conspicuous (Fig. 65).
Sucrose nitrate agar: Colony usually flat,
level with agar surface; concentric zonation
distinct or absent. Surface glossy or powdery.
Color usually white, sometimes pinkish-
buff or cream-buff. Sporangia absent to
fairly abundant, always formed some dis-
tance above the surface of the agar. In some
cultures, coils form which break up into
spores as in Streptomyces.
Glucose-asparagine agar: Growth poor,
shghtly elevated and minutely ridged, slop-
ing to the fimbriate margin. Surface of
central region powdery with white aerial
hyphae. Sporangia absent.
Potato-glucose agar: Growth usually good,
center elevated with irregular bumps and
ridges; margin flat and even with surface of
agar. Color of colony at first creamy, be-
coming tawny and then brown, after which
white hyphae appear,
usually spreading to cover the entire culture.
Sporangia usually formed in vast numbers,
the white areas becoming rosy pink as the
sporangia mature; the pinkish areas are
frequently minutely pocked. Surface moist
at first, appearing dry and floccose as aerial
hyphae and sporangia are formed. Agar
floccose spots of
colored reddish-brown with a vinaceous
tinge.
Nutrient agar: Growth fair, color usually
cream-buff, rarely buff-brown. Surface usu-
ally glossy, sometimes powdery with aerial
hyphae which may be united to form many
upright fascicles. Sporangia absent.
Habitat: Garden soil in North Carolina
and forest litter in Holland and Denmark
(Van Brummelen and Went, 1957).
Remarks: Nonomura and Ohara (1960b)
found the genus Streptosporangium widely
distributed in the soils of Japan. In addition
to the original S. roseum Couch, four new
species were isolated: S. album, S. viridial-
bum, S. amethystogenes, and S. vulgare. These
were classified on the basis of the color of
the aerial mycelium and formation of soluble
pigment on oatmeal agar media.
Chapter
Incompletely
13
Described Species
of Actinomycetes
Numerous isolates of cultures of actino-
mycetes listed in the literature, under a
variety of different
names, could not be identified at present,
generic and specific
due to insufficient descriptions. They are
reported here as “incompletely described.”
The naming of such cultures was based either
on a casual observation or on the assumed
occurrence of such an organism in a certain
disease condition. Frequently the particular
organism was not even obtained in pure
culture, but was given
‘
c
1 name, often for
the mere purpose of obtaining for the author
credit for the particular isolation or observa-
tion. In other cases, it is fairly certain that
the culture said to have been isolated from
a disease condition was nothing more than a
dust contamination.
In the preparation of this list, the author
has used freely the carefully collected records
of Brumpt (1939), Dodge (1935), Baldacci
(1944), and Krassilnikov (1949). Very few
of these descriptions were complete enough
to include the recorded cultures among the
readily identifiable species. This is partic-
ularly true, even in recent years, where
media of unknown composition were used
for descriptive purposes.
Not all the synonyms are recorded here.
For further detail, the reader is referred to
the reviews of Chalmers and Christopherson
(1916), and Dodge (1935),* especially for the
*Some of these compilations may be designated,
quite properly, in the words of Erikson (1940), as
315
pathogenic or largely would-be pathogenic
and to Brumpt (19389) and
Krassiinikov (1949) for the nonpathogenic
forms. Only those synonyms that would
tend to throw light upon the systematic
position of the culture are listed here.
Since the name
organisms,
ce
Actinomyces” has been
largely used for the incompletely described
cultures, it 1s left as such, and the list is
recorded in the order of species. Where the
name Nocardia was originally used, it is
either reported as a synonym of ‘‘Actino-
myces,”’ or under Nocardia. The same prin-
ciple was applied to JM/icromonospora and
other well-recognized genera.
No serious attempt was made to record
all the other listings of cultures believed to
be actinomycetes, especially those that have
been insufficiently described under a variety
of different Cladothrix,
Discomyces, etc. Most of these names appear
«
c
names, such as
to be synonyms of those listed above. Only a
few of the names listed under Streptothria
and Oospora are included under the ‘‘Ac-
tinomyces.”’ No effort was made to list cul-
tures for which only a generic name was
given without any specific designation, or
which were recorded by a number only.
A large number of species have recently
been listed under the genus Streptomyces
without any description at all or with a
totally insufficient description. Irequently
“veritable mausoleums wherein the errors of the
past are indiscriminately embalmed.”’
316
the description is credited to a company and
not to an individual scientist. This was
done primarily as an effort to establish
priority for an antibiotic isolated from such
a culture, or for patent purposes. These
designations are placed in a separate cate-
gory, with emphasis on the antibiotic. A
group of cultures described by Gause ef al.
(1957) has also been placed in a separate
category, since no decision can be reached
as yet concerning their synonymity with
previously described species.
For more complete lists of names of
actinomycetes, comprising both genera and
species, the reader is referred to Buchanan
and Lessel (1959), Lessel (1960), and es-
pecially to their forthcoming treatise ‘Index
Bergeyana.”’
According to Buchanan and Lessel (1959),
there are now about 3000 names given to
different strains of bacteria that are recog-
nized as belonging to one or another of the
genera of the order Actinomycetales. They
emphasize, however, that ‘‘this is not to be
interpreted as meaning that there are three
thousand species, for a large proportion of the
names (probably two-thirds) are notavailable
for use because they were not validly pub-
lished, or are homonyms, or synonyms, or
are not binary combinations, or were pro-
posed as hypothetical names, or were in-
sufficiently described and are to be regarded
as naked names (nomina nuda) or as
doubtful
officially
illegitimate as
names (nomina dubia) or are
beeause
other
rejected names, or
contravening some
nomenclatural rule.”
An attempt has been made to present in
the following lists some of the incompletely
Many
additional names are found in the previous
described forms of actinomycetes.
chapters as synonyms of well described
organisms.
The first list comprises the forms described
under the names Actinomyces (A), Strepto-
thrix (St.), or Oospora (O).
THE ACTINOMYCETES, Vol. II
The name Actinomyces used in this list is
not to be confused with the genus Actino-
myces recognized at present, although some
of the cultures so designated here would no
doubt be considered as members of this
genus.
A. acidoresistans, a culture obtained from
Pribram collection in Vienna. IMRU
3049.
A. actinoides (Smith, 1918) Bergey, 1923.
A. actinomorphus (Gray and Thornton,
1928) Bergey, 1930.
St. actinomyces Rossi-Doria, 1891.
St. actinomycotica Foulerton, 1899.
A. aerugineus Wollenweber, 1920.
A. agrestis (Gray and Thornton,
Bergey, 1930.
St. albtdo Chester, 1901.
A. albidoflava (Rossi-Doria, 1891) Ford, 1927
. albidofuscus Berestnew, 1897.
. albido-fuscus Neukireh, 1902.
. alboatrus Waksman and Curtis, 1916.
. albopurpureus Duché, 1934.
. albus-acidus Neukirch, 1902.
. albus var. actdus Nannizzi Pollacei, 1934.
. albus var. alfa Ciferri, 1927.
. albus asporogenes Berestnew, 1897.
. albus chlamydosporus Wrassilnikov, 1949.
1928)
‘ 2 a ee i
a—~ Se Se ee oe oe oe oe
»
i. K
K
. albus vulgaris IKrassilnikoyv, 1941.
St. alpha Price-Jones, 1900.
1. allenbachi Sartory and Meyer, 1932.
A. almquisti Duché, 1934.
1. americanus (Cohnistreptothrix americana
Chalmers and Christopherson, 1916)
Dodge, 1935.
A. anaerobies (Plaut, 1920; Oospora anae-
robies Sartory, 1923) Dodge, 1935.
A. anaerobicus (Plaut, 1920) Ford, 1927.
St. aquatilis Johan-Olsen, 1893.
A. aquatilis Salimowokaja, 1928.
nikov states that it is related to A. glaucus.
A. arborescens (Edington, 1887) Gasperini,
1894.
A. aromaticus IwrassilInikoyv, 1941.
Sartory
Ixvassil-
A. asteroides serratus and
Meyer, 1930.
var.
INCOMPLETELY DESCRIBED SPECIES OF ACTINOMYCETES 31
A. atypica pseudoturberculosa Hamm and
Keller, 1909.
St. aurea (Saint-Sévérin, 1895) Ford, 1927.
A. aureus Lachner-Sandoval, 1898.
Syn. N. aurea Castellani and Chalmers,
1913.
A. avadi Dodge, 1935.
A. baarnensis Duché, 1934.
Syn. of A. viridis Duché, 1934.
A. bahiensis (daSilva, 1919) Brumpt, 1927.
A. bellisari Dodge, 1935.
A. berestneffi (Chalmers and Christopherson,
1916) Brumpt, 1939.
Syn. NV. berestneffi Chalmers and Christo-
pherson, 1916.
A. berardinisi (Namyslowsky, 1912) Brumpt,
1939.
St. beta Price-Jones, 1900.
A. bicolor Trolldenier, 1903.
Syn. N. bicolor deMello and Fernandes,
1919.
A. bolognesii-chiurcot
1935.
A. bostroemt Baldacei, 1937.
A. bovis albus Gasperini, 1894.
St. bovis communis Foulerton
Jones, 1901.
A. bovis farcinicus Gasperini, 1894.
(Vuillemin) Dodge,
and Price-
A. bovis luteoroseus Gasperini, 1894.
A. bovis sulfureus Gasperini, 1894.
A. bovis var. nigerianus Erikson, 1935.
A. bronchialis (Sartory and Lasseur, 1914)
Brumpt, 1959.
A. bronchiticus
Brumpt, 1939.
Svn. Anaeromyces bronchitica Castellani
et al., 1921.
A. brumpti (Bordjoski and Milochevitch,
1935) Brumpt, 1939.
A. bruni (Chalmers and Christopherson,
1916) Brumpt, 1939.
Syn. V. brunt Chalmers and Christopher-
son, 1916.
A. buccalis (Roger et al., 1909) Brumpt,
1939.
(Castellani et al., 1921)
“J
Syn. O. buccalis Roger et al., 1909.
N. buccalis Castellani and Chalmers,
1913.
A. cameli (Mason, 1919) Ford, 1927.
A. caminiti Ford, 1927.
St. candida (Petruschky, 1898) Ford, 1927.
Syn. V. candida Castellani and Chalmers,
1913.
A. canis (Rabe, 1888) Gasperini, 1894.
A. canis familiaris Rivolta, 1884.
St. caprae (Silberschmidt, 1899) Ford, 1927.
A. carnea (Rossi-Doria, 1891; Gasperini,
1894; Kruse, 1896) Ford, 1927.
Syn. N. carnea Castellani and Chalmers,
1913.
A. carougeaw (Gougerot,
1939.
Syn. NV. carougeaw Castellani and Chalm-
ers, 1913.
A. catarrhalis (Bailly, 1921) Brumpt, 1939.
A. cati (Rivolta, 1878) Gasperini, 1894.
A. caviae (Snijders, 1924) Erikson, 1935.
1910)
1909) Brumpt,
A. cerebriformis
Brumpt, 1939.
A. cereus Lieske, 1921.
A. chalmersi (deMello and Fernandes, 1919)
Dodge, 1935.
Syn. V. chalmersi deMello and Fernandes,
1919.
A. christophersoni (deMello and Fernandes,
1919) Dodge, 1935.
Syn. NN. christophersoni
Fernandes, 1919.
A. chromogenes (Gasperini,
1896) Ford, 1927.
O. chromogenes Lehmann and
1896.
A. cinereo-niger Lieske, 1921.
St. cinereonigeraromaticus (Berestnew, 1897)
Neukirch, 1902.
A. citrea (Gasperini, 1894) Ford, 1927.
A. citrotremeus Nannizzi Pollacci, 1934.
A. cloacae Brussoff, 1919.
A.
A.
AN
(Namyslowsky,
deMello and
1891; Kruse,
Neumann,
coccocidus Krassilnikov, 1955.
colorata Sanborn, 1926.
congolensis (Baerts, 1925) Brumpt, 1939.
318
A. convolutus (Chalmers and Christopherson,
1916) Brumpt, 1939.
Syn. N. convoluta Chalmers and Christo-
pherson. According to Gonzalez
Ochoa and Sandoval (1956), this is a
synonym of NV. asteroides.
»
—
4. coremiales Wrassilnikov, 1949.
A. cretaceus (KKriiger, 1905) Wollenweber,
1920, who considered it as a variety of A.
albus.
Syn. O. cretacea WKriiger, 1905.
A. cruoris (Macfie and Ingram,
Brumpt, 1927.
Syn. N. cruoris Macfie and Ingram, 1921.
A. crystallophagus (Gray and Thornton,
1928) Bergey, 1930.
A. cuniculi (Schmorl) Gasperini, 1894.
Svn. N. cuniculi deMello and Fernandes,
1919.
St. cuniculi Foulerton and Price-Jones, 1901.
A. cylindraceus (dekkorté, 1918) Brumpt,
1939.
Syn. N. cylindracea delxorté, 1918.
O. cylindracea Sartory, 1923.
A. dassonvillet (Broeg - Rousseu,
Brumpt, 1959.
Syn. NV. dassonviller Liégard and Landrieu,
1911.
A. decussatus (Langeron
1912) Brumpt, 1939.
Syn. Discomyces decussatus Langeron and
Chevalier, 1912
N. decussata Castellani and Chalm-
ers, 1913.
O. decussata Sartory, 1923.
A. denitrificans (Nikolaeva, 1914) Krassil-
nikov, 1949.
A. dermatonomus
1921)
1907)
and Chevallier,
Bull, 1929
Thompson
(Polysepta
dermatonomus and Bisset,
1957).
A. dispar (Vidal, 1882) Brumpt, 1939.
A. donnae Dodge, 1935.
A. dort (deBeurmann and Gougerot, 1906)
Brumpt, 1939.
O. doriae Sauvageau and Radais, 1892.
THE ACTINOMYCETES, Vol. II
A. egyptt Gohar et al., 1954 (Ettlinger et al.,
1958).
A. elaeagni Roberg, 1934. Produces tubers
on the roots of the oleander plant.
A. elephantis primigenit Omeliansky, 1909.
A. enteritidis (Pottien, 1902) Brumpt, 1927.
Syn. N. enteritidis Castellani and Chalmers,
1913.
St. eppingert (Rossi-Doria,
slowsky, 1912.
A. equi (Chalmers and
1916) Brumpt, 1939.
Syn. NV. equi (Dean, 1900) Chalmers and
Christopherson, 1916.
A. erystpeloides (Neumann and Lehmann,
1895) Lachner-Sandoval, 1898.
Syn. N. rosenbachi (Gougerot, 1913).
St. erythrea Foulerton, 1902.
St. farcinica (Trevisan, 1889)
1891.
A. ferrugineus (Naunyn) Krassilnikov, 1949.
1891) Namy-
Christopherson,
tossi-Doria,
Syn. N. ferruginea de Toni and Trevisan.
A. flava (Sanfelice, 1904) Ford, 1927.
A. flavus (Chester, 1901) Dodge, 1935.
A. fluorescens KKrassilnikov, 1955.
St. Foerster? Cohn, 1875. Observed in con-
cretions in the lachrymal canal. A faculta-
tive anaerobe, not pathogenic for labora-
tory animals. Numerous synonyms. of
this name are found in the literature. It is
sufficient to lst Leptothrix oculorum
Sorokin, 1881; Cl. foersteri Winter, 1884;
Cl. dichotoma Macé, 1888; N.. forstera
Trevisan, 1889; O. foérsteri Sauvageau et
Radais, 1892; St. fdrsterz Kruse, 1896;
N. aurea Saint-Sévérin, 1902; Cohn-
istreptothrix silberschmidtt. Chalmers and
Christopherson, 1916.
A. foulertont (Chalmers and Christopherson,
1916) Brumpt, 1939.
Syn. NV. foulertont Chalmers and Christo-
pherson, 1916.
St. freer? Musgrave and Clegg, 1907.
Syn. A. freer: Bergey, 1923.
Pr. freer? WKrassilnikov, 1949. Defi-
nitely a Nocardia.
INCOMPLETELY DESCRIBED SPECIES OF ACTINOMYCETES
A. fusca S6hngen and Fol, 1914.
A. fuscus (Karwacki, 1911) Brumpt, 1939.
Syn. N. fusca Castellani and Chalmers,
1913.
A. gabritschevski (Berestnew, 1898) Krassil-
nikov, 1941.
A. gartent Brumpt, 1927.
Syn. NV. gartent Gougerot, 1913.
A. gedanensis (LOhlein, 1909) Bergey, 1923.
A. gedanensis (Scheele and Petruschky,
1897) Ford, 1927.
Syn. N. gedanensis Chalmers and Christo-
pherson, 1916.
St. gelatinosus Johan-Olsen, 1897.
A. genesti (Frées, 1930) Dodge, 1935.
Syn. N. genesiz Frées, 1930.
A. gibsoni Dodge, 1935.
A. goensis (deMello and Fernandes, 1919)
Dodge, 1935.
Syn. N. goensis deMello and Fernandes,
1919.
A. gonadiformis Bergey, 1930.
A. graminearus (Berestnew, 1897) IXrassil-
nikov, 1949.
A. graminis (Bostroem, 1891) Topley and
Wilson, 1929, 1946.
A. griseo-viridis Nikolaeva, 1914.
A. griseus variabilis Krassilnikov, 1949.
A. grubert (Terni, 1894) Sanfelice, 1904.
Syn. N. grubert Blanchard (In Bouchard,
1896).
A. gueguent (Brumpt, 1921) Brumpt, 1939.
Syn. D. lingualis Guegen, 1908.
N. lingualis Castellani and Chalmers,
1913.
A. guerrai (Langeron, 1929) Brumpt, 1939.
A. guignardi (Sauvageau and Radais, 1892)
Ford, 1927.
A. gypsoides Henrici and Gardner, 1921.
A. halotricus ZoBell and Upham, 1944.
A. heimi Duché, 1934.
A. hobnesi (Gedoelst, 1902) Nannizzi Pollacci,
1934.
A. hofmanni
1894.
A. hominis Bostroem, 1890.
(Gruber, 1891) Gasperini,
319
St. hominis II, III, IV (Foulerton, 1906
1910).
St. hominis (Berestnew, 1897).
Not A. hominis Waksman, 1919.
St. humifica Johan-Olsen, 1897.
A. incanescens Wollenweber, 1922.
A. indicus (Kanthack, 1893) Brumpt, 1939
Syn. N.
A. innominatus Baldacci, 1939.
A. interproximalis (Fennel, 1918) Ford, 1927.
A. tnvulnerabilis (Acosta and Rossi, 1893;
Kruse, 1896; Lachner-Sandoval, 1898)
Ford, 1927. Isolated as laboratory con-
tamination, and also from water. With-
stands heating at 100—-120°C. Grows in
media containing 0.5 per cent copper sul-
fate, 0.5 per cent phenol, 1 per cent boric
acid, and 0.01 per cent mercuric chloride.
St. vsraeli Kruse, 1896.
A. japonica (Petruschky, 1913) Ford, 1927.
A. japonicus (Aoyama and Miyamoto, 1901)
Brumpt, 1959.
A. jolly (Vuillemin, 1920) Brumpt, 1927.
A. keratolyticus (Acton and McGuire, 1931)
Brumpt, 1939.
A. krainskii Duché, 1954.
A.krauser (Chester, 1901) Brumpt, 1927;
Ford, 1927.
Syn. NV. krauset Chalmers and Christopher-
son, 1916.
A. lacertae (Terni, 1896) Foulerton, 1912.
Syn. A. sulfureus lacertae Berestnew, 1897.
A. lanfranchii Sani, 1916.
indica var. flava Kanthack, 1893
Syn. N. lanfranchii deMello and Pais,
1918.
A. lasseret (Verdun, 1912) Brumpt, 1939.
Syn. N. lasserer (Verdun) Castellani and
Chalmers, 1913.
St. lathridi (Petruschky, 1898) Ford, 1927.
A. leishmani (Chalmers and Christopherson,
1916) Brumpt, 1939.
A. lepromatis (deSouza-Araujo,
Brumpt, 1939.
St. leucea Foulerton, 1902.
St. leucea saprophytica Foulerton, 1912.
A. levyi Dodge, 1935.
1929)
320
A. liesket Duché, 1934.
Actinobacillus — ligniersi
Brumpt, 1939.
A. liguire (Urizer, 1904) Nannizzi Pollacci,
1934.
A. lingualis (Weibel, 1888; non Gueguen,
1908) Brumpt, 1939.
A. liquefaciens (Hesse, 1892) Brumpt, 1939.
Syn. N. liquefaciens Castellani and Chal-
mers, 1913.
A. liquefaciens (Garten, 1895) Ford, 1927.
A. londinensis (Chalmers and Christopher-
son, 1916) Brumpt, 1939.
Syn. N. londinensis Chalmers and Christo-
pherson, 1916.
4. longisporus Krassilnikov, 1941.
A. longisporus ruber Krassilnikov, 1941.
A. longisporus griseus Kkrassilnikov, 1941.
4
1
(Brumpt, 1910)
. longissimus Krassilnikov, 1941.
luteolus (Foulerton
Brumpt, 1939.
A. luteo-roseus Gasperini, 1894.
A. macrodipodidarum (Fox, 1923) Dodge,
1935.
Syn. N.
4. malencont Duché, 1934.
A. matruchoti (Mendel, 1919) Brumpt, 1939.
{
1
and Jones, 1910)
macrodipodidara Fox, 1923.
. melanogenes Rubentschik, 1928.
4. melanoroseus Issatschenko, 1927.
A. melanosporeus IKrainsky, 1914.
St. melanotica Price-Jones, 1903.
A. metchnikovi (Sauvageau and
1892) Ford, 1927.
A. mexicanus (Boyd and Crutchfield, 1921)
Brumpt, 1939.
Syn. N. mexicana Ota, 1928.
A. micetomae (Greco, 1916) Dodge, 1935.
Radais,
Syn. O. micetomae Sartory, 1923.
4. mshagiensis Salimowskaja, 1928.
A. microflavus Krainsky, 1914.
{. mihi Caminiti, 1907.
1. mineaceus (Ixruse, 1896) Lachner-
Sandoval, 1898.
A. minimus (LeCalvé and Malherbe, 1900)
Dodge, 1935.
A. minutissimus (Burchard) Brumpt, 1927.
THE ACTINOMYCETES, Vol. II
Syn. N. minutissima (Verdun, 1912) Cas-
tellani and Chalmers, 1913.
A. mucosus Basu, 1943.
A. multifidus Krassilnikov, 1941.
A. muris-ratti (Schottmiller, 1914) Ford,
1927.
A. musculorum Hertwig, 1886.
A. mutabilis Masumoto, 1948; Gause et al.,
1957.
A. myricae Peklo, 1910.
St. necrophora Wilhelm, 1902.
A. necrophorus (Fligge, 1886) Lehmann and
Neumann, 1926.
A. neddeni (Namyslowsky, 1912) Brumpt,
1939.
A. neschezadimenki (Chalmers and Christo-
pherson, 1916) Dodge, 1935.
A. nmicollet (Delanoé, 1928) Brumpt, 1939.
Syn. N. nicollez Delanoé, 1928.
A. mger (Rossi-Doria, 1891) Krassilnikoy,
1949.
A. niger aromaticus Berestnew, 1897.
St. nigra (Rossi-Doria, 1891) Sanfelice, 1904.
A. nigricans Willian and Fehér, 1935.
A. nigrificans (IKkriger, 1905) Wollenweber,
1920.
St. nigrescens Foulerton, 1901.
A. nitrogenes Sartory et al., 1936.
A. nocardi (Foulerton, 1901)
1911.
A. nodosus (Beveridge, 1941) Hagan, 1948.
A. nondiastaticus Bergey, 1919. Grows at
65°C; differs from S. thermodiastaticus in
not decomposing starch.
A. non-fluorescens Krassilnikov, 1955.
A. ochraceus Neukirch, 1902.
A. ochroleucus Neukirch, 1902. A culture
described under same name was isolated
from diseased potatoes by Wollenweber
(1922) who considered it as a variety of
A. albus.
A. odoratus Wrassilnikov, 1941.
Buchanan,
A. odorifer Woelz, 1936.
St. oidioformis Johan-Olsen, 1893.
A. oligocarbophilus (Beijerinck
Delden, 1903) Lantzsch, 1922.
and Van
INCOMPLETELY DESCRIBED SPECIES OF ACTINOMYCETES 3:
St. orangica Berestnew, 1897.
A. orangico-niger Lieske, 1921.
A. orangicus (Rossi-Doria) Lieske, 1921.
A. panginensis (deMello and Fernandes
1919) Dodge, 1935.
Syn. NV. panginensis deMello and Fernan-
des, 1919.
St. paulotrophus Beijerinck, 1914.
A. pelogenes Sawjalow, 1913.
A. penicilloides Sartory and Meyer, 1936.
A. phagocidus WKrassilnikov, 1955.
A. phenotolerans Werkman and Patrick, 1932.
Isolated from granuloma in man. Grows
well in phenol-containing media. Gon-
zalez Ochoa and Sandoval (1956)
sidered it a synonym of N.
A. pijpert (Castellani and Chalmers, 1919)
Brumpt, 1939.
Syn. N. pijypert Castellani and Chalmers,
1919.
A. pinoyt (deMello and Fernandes, 1919)
Dodge, 1935.
Syn. N. pinoyz deMello and Fernandes,
1919.
A. plurichromogenus (Caminiti, 1907) Dodge,
1935.
Svn. N. plurichromogena Caminiti, 1907.
A. pluricolor Gasperini, 1894.
Syn. N. pluricolor Terni, 1894; N. pluri-
color deMello and Fernandes, 1919.
A. pluricolor diffundens (Berestnew, 1897)
Lieske, 1921.
A. polychromogenus Dodge, 1935.
A. ponceti (Verdun, 1912) Brumpt, 1939.
Syn. N. ponceti (Verdun, 1912) Castellani
and Chalmers, 1919.
con-
asteroides.
A. pretorianus (Pijper and Pullinger, 1927)
Brumpt, 1939.
Syn. NV. pretoriana (Pijper and Pullinger,
1927).
A. protea (Schtirmayer, 1900) Ford, 1927.
A. pseudonecrophorus Harris and Brown,
1927.
A. pseudotuberculosae
Brumpt, 1939.
A. pseudotuberculosis (Keller) Dodge, 1935.
(Flexner, 1S98)
bo
—
Syn. JN.
Fernandes, 1919.
A. pseudotuberculosus (Flexner, 1898) Leh-
mann and Neumann, 1912.
A. pulmonalis Burnett, 1909; (Roger et al.,
1909) Brumpt, 1939.
A. puntonii Lopez Ortega, 1934.
A. purpurogenus Waksman and Curtis, 1916.
A. purpureus (Orloff, 1913) Brumpt, 1939.
St. putori Dick and Tunnicliff, 1918.
A. putridogenes (Vezspremi, 1907) Nannizzi
Pollaeci, 1934.
A. putrificus Nikolaeva, 1914.
A. pyocyaneus Rullmann, 1895.
St. pyogenes Caminiti, 1907.
A. pyogenes (Chalmers and Christopherson,
1916) Dodge, 1935.
A. radiatus (Namyslowsky, 1912) Brumpt,
1939.
St. ratt? Schottmiller, 1914.
A. ribeyrot Dodge, 1935.
A. rivieret (Verdun, 1912) Brumpt, 1939.
A. rodellae Dodge, 1935.
A. rogersii Brumpt, 1939.
Syn. N. rogersi deMello, 1919.
A. rosaceus Lieske, 1921.
O. rosella Kriiger, 1905.
A. rosenbachi (IKkruse, 1896) Holland, 1920.
A. roseolus Nadson, 1903.
O. rubea Wilbert, 1908.
St. rubea Chalmers and Christopherson, 1916.
A. ruber (Ikruse, 1896) Santelice, 1904.
St. rubra (Kruse, 1896) Ford, 1927.
A. rubidaureus Lachner-Sandoval, 1898.
Syn. A. mordoré Thiry, 1897.
pseudotuberculosis deMello and
O. mordoré Sartory, 1923.
N. thiryer deMello and Pais, 1918.
A. sabrazes (Ferre and Faguet, 1895) Dodge,
1935.
A. saharae Willian and Fehér, 1935.
A. salvati Langeron, 1922; Fontoynont and
Salvat, 1922.
A. sanfelicer (Redaelli,
Pollaeei, 1934.
Syn. NV. sanfelicez Redaelli, 1928.
A. sanguinis Basu, 1937.
1928) Nannizzi
o22 THE ACTINOMYCETES, Vol. II
St. sanninit Ciferri, 1922.
A. saprophyticus Gasperini, 1892. Lieske,
1921.
A. saprophyticus var. chromogenes Gasperini,
1892.
A. sartoryt Dodge, 1935.
A. scabies var. anglica Baldacci and Spalla,
1956.
A. sendaiensis
Brumpt, 1939.
A. septicus MacNeal and Blevins, 1945.
A. serratus (Sartory et al., 1930) Dodge, 1936.
A. silberschmidti (Chalmers and Christo-
pherson, 1916) Dodge, 1936.
(Ping-Ting-Huang, 1933)
Syn. NN. silberschmidti deMello and
Fernandes, 1919.
A. somaliensis (Brumpt, 1906) St. John-
Brooks, 1931.
A. sommert (Greco, 1910) Brumpt, 1939.
A. spinae Velich, 1929.
A. spinosporus (Spini) Verlich, 1914.
St. spirilloides Johan-Olsen, 1893.
A. spitzi (Ligniéres and Spitz, 1904) Dodge,
1935.
Syn. O. spitzi Sartory, 19
A. splenicus (Gibson, 1930) Brumpt, 1939.
Syn. NV. splenica Gibson, 1930.
A. spumalis (Sartory, 1923) Dodge, 1935.
Appears to be a Nocardia.
St. taraxert cepapit (Schottmiller,
Ford, 1927.
A. tarozzii (Miescher, 1917) Dodge, 1935.
St. tartarz Sanfelice, 1904.
O. tenax KKriiger, 1905.
A. tenuis (Castellani, 1911) Dodge, 1935.
Syn. N. tenuis Castellani, 1911.
S. termitum Duché et al., 1951.
A. thermotolerans Lieske, 1921.
A. thibierget (Ravaut
Brumpt, 1939.
Syn. NV. thibierget Castellani and Chalmers,
1913.
A. thjottae (Thjetta and Gundersen, 1925)
Dodge, 1935.
oF
2
ao.
1914)
and Pinoy, 1909)
A. thwilliert (deToni and Trevisan, 1889)
Brumpt, 1939.
Syn. N. thwalliert Vuillemin, 1931.
4. tossicus (Rossi, 1905) Dodge, 1935.
A. totschidlowskii Serbinov, 1925.
A. toxicus IKrassilnikov, 1955.
A. transvalensis (Pijper and Pullinger, 1927)
Brumpt, 1939.
4. tricolor Wollenweber, 1922.
4. tyrosinaticus Beijerinck, 1914.
A. urethritidis (Rocek, 1920) Brumpt, 1939.
1. urinarius (Pijper, 1918) Brumpt, 1939.
4. valvulae (deMello and Pais, 1918) Nan-
nizzi Pollacei, 1934.
Syn. N. valvulae deMello and Pais, 1918.
A. valvularis (Luginger, 1904) Ford, 1927.
A. valvulas destruens bovis Luginger, 1904.
A. verrucosus Nadson 1903; (Miescher, 1917)
Brumpt, 1939.
A. violacea (Rossi-Doria, 1891) Ford, 1927.
A. waksmanii Bergey, 1930.
A. xanthostromus Wollenweber, 1922.
St. zopfi (Casagrandi) Caminiti, 1907.
The above is only a partial list of incom-
pletely described cultures, most of which
would now be included in the genus Strepto-
Some cultures were listed as Dzs-
comyces, pleuriticus Vachetta
(1882) and D. pleuritticus canis familiaris
Rivolta. Various others were given under
the generic names Bacillus, such as B.
actinoides Smith, 1918; Bacterium, such as
B. actinocladothrix Afanassiev, 1888; J/yco-
bacterium, such as M. paraffinicum (Davis
et al., 1956), also under the anaerobic genus
Actinobacterium, including A. meyert, A.
abscensus, A. cellulitis (Linhard, 1949); and
Act. ligniersi
myces.
such as D.
as Actinobacillus, such as
Brumpt, 1910.
A large number of cultures were simply
given numbers, as done by Drechsler,
Lieske, and many others. Finally, some were
mentioned under a generic name, without
even the aflixation of a number, such as
Actinomyces sp., Streptomyces sp., ete. All of
these, with very few exceptions, need not be
considered any further here.
Many of the above cultures have been
isolated from excretions of man and labora-
INCOMPLETELY
tory animals. Some were found to be
pathogenic to experimental animals. Most
of them were not tested, however, for their
pathogenicity.
Organisms Belonging to the Genus No-
cardia
In addition to many of the forms listed
previously, certain other incompletely de-
scribed forms which probably belong to the
genus Nocardia may be mentioned. These
have been described under the generic names
of Nocardia (N.), (As.),
Proactinomyces (Pr.).
Asteroides and
N. actinomyces Trevisan, 1889.
N. albida Chalmers Christopherson,
1916.
N. albosporea Chalmers and Christopherson,
1916.
N. appendicis Chalmers and Christopherson,
1916.
Pr. aquosus Turfitt, 1944.
Pr. asteroides var. crateriformis
1937.
Pr. asteroides var. decolor Baldacei, 1937.
N. bifida (B. bifidus Tissier, 1901).
N. bovis Gougerot et al., 1934.
N. cunicult Snijders, 1924.
Pr. cyaneus (Beijerinck, 1914) Krassilnikov,
1941.
Pr. cyaneus antibioticus Gause, 1946.
N. erythropolis (Gray and Thornton, 1928)
Waksman and Henrici, 1948.
N. filiformis (Boas, 1897) Vuillemin, 1931.
N. krainskii Chalmers and Christopherson,
1917.
N. lignieresi (Brumpt, 1910) Chalmers and
Christopherson, 1916.
As. liskeyi Puntoni and Leonardi, 1935.
and
Baldacci,
N. minima (Jensen, 1931) Waksman and
Henrici, 1948.
N. pluricolor Namyslowsky, 1912.
As. pseudocarneus Puntoni and Leonardi,
O35:
Pr. pseudomadurae Baldacci, 1945.
N. ramosa (B. ramosus Veillon and Zuber,
1898).
DESCRIBED SPECIES OF ACTINOMYCETES oo
Pr. restrictus Turfitt, 1944.
N. ripens (Eklund, 1883) Vuillemin, 1931.
N. saprophytica Chalmers and Christopher-
son, 1916:
N. sylvodorifera Castellani, 1911.
Various other specific names for organisms
probably belonging to the genus Nocardia
have been listed under several other genera,
such as Cladothrix (Cl. actinomyces Rossi-
Doria, 1891; Macé, 1897); Cohnistreptothrix
(Co. americana Chalmers and Christopherson,
1916); Discomyces (D. asteroides Eppinger,
1891; Godoelst, 1902); Flavobacterium (F.
salmonicolor den 1927;
Bergey, 19380); A/ycobactervum (AM. albuvia-
Dooren de Jong,
lum Bergey, 1923); Serratia (S. corralina
Hefferan, 1904; Bergey, 1923), and others.
Krassilnikov (1949) listed or
numerous other forms belonging to this
genus under the names of Nocardia, Proac-
tinomyces, Mycobacterium, Bacillus, Bac-
terium, Brevistreptothrix, Cladothrix, Cohn-
deseribed
istreptothrix, Discomyces, and various others.
Thompson and Bisset (1957) suggested a
new generic name Polysepta.
Descriptions Incomplete or Needed for
Actinomycetes Preducing Specific
Antibiotics and Vitamins
Because of the growing importance of
actinomycetes as producers of antibiotics
and because of the desire to claim priority
for a new antibiotic, many names have been
introduced for species of Streptomyces and
Nocardia. Often these names are mentioned,
with incomplete descriptions or with no
descriptions, in the patent literature or even
in trade journals. An attempt was made to
collect these. The list (Table 27) is far from
complete, however.
A number of new species of actinomycetes
belonging to the genus Streptomyces have
been created by Gause et al. (1957-1959),
who justified this by the fact that there was
a great need for describing more organisms
capable of producing antibiotics. Only a
limited attempt was made in these descrip-
Ne}
Ne
THE ACTINOMYCETES, Vol. 11
TABLE 27
Incompletely described antibiotic-producing species of Nocardia and Streptomyces
Organism Reference Antibiotic or vitamin
Nocardia
N. acidophilus J. Bacteriol. 54: 281, 1947 Mycomycin
Pr. actinoides Antibiotiki 2(5): 44, 1957 Actinoidin
N. lurida Antibiotics Ann. 1956-1957, 687, 693, 699 Ristocetin
N. narasinoensis J. Antibiotics (Japan) 7A: 1, 1954; 8B: 253, 1955 Nocardorubin
Streptomyces
S. albicans Med. Parasitol. USSR 4, 1947 Actinolysin
S. albidofuscus J. Antibiotics (Japan) 6A: 140, 1953 Pyridomycin
S. albulus Abstr. Papers 134th Ann. Meet. Am. Chem. Soc. Antitumor substances
Chicago, 1958, 22.
S. aminophilus Antibioties Ann. 1955-56, 236 Antibiotic 1968
S. arabicus J. Antibiotics (Japan) 9B: 62, 1956 Croceomycin
S. bacillaris Folia Biol. 4: 260, 1958 Various antibiotics
S. badius J. Antibiotics (Japan) 3: 582, 1949 Streptothricin
S. blastmyceticus J. Antibioties (Japan) LOA: 40, 1957 Blastmycin
S. caiusiae J. Sei. Ind. Res. India 1l6e: 76-81, 1957 Antibiotic X
S. carcinomyceticus Chemotherapy (Tokyo) 3: 129, 1955 Carcinomycin
S. cellostaticus Tohoku J. Exptl. Med. 67: 173, 1958. Cellostatin
S. chatlanoogensis Antibiotics & Chemotherapy 9: 398, 1959 Tennecetin
S. chibaensis J. Antibiotics (Japan) I1A:81, 1958 Cellocidin
S. chrestomyceticus Giorn. Microbiol. 7: 242, 1959 Aminocidin
S. cinnamonensis Ann. Soc. Biol. Pernambuco 13: 3, 1955; 14: 9, Streptothricin-like
1956 antibiotic
A. circulatus var. Antibiotiki 5(4): 3, 1960. Monomycin
MONOMYCUNL
A. coeruleus antibio- Antibiotiki 2(1): 25, 1957 An antibiotic
licus
S. colombiensis U.S. 2,595,499, May 6, 1952 Vitamin By»
S. ehimensis J. Antibiotics (Japan) 7B: 168, 1954 Candimyein
S. fasciculatus Antibiotics & Chemotherapy 3: 718, 1953 Amicetin
S. flavofungini Nature 181: 908, 1958 Flavofungin
S. fluorescens Folia Biol. 4: 259, 1958 Fluorin
A. fluorescens Antibiotiki 5(1): 25, 1960 Actinomycin
A. fradiae var. spiralis Antibiotiki Ll: 4, 1956 Colimyein; probably
neomycin
S. ganmycicus J. Antibiotics (Japan) 9A: 8, 113, 1956; 9B: 160, Ganmycin + Carcino-
1956 mycin
S. ganmyceticus J. Antibiotics (Japan) 9A: 6, 9, 113, 1956 Carzinocidin
S. globisporus tundra- Bull. Moscow Soc. Nat. Sci. Biol. 62(2): 79, 1957. Tundramycin
mycint
S. graminofaciens Antibiotics & Chemotherapy 3: 1288, 1953; Streptogramin
Antibioties Ann. 1953-1954, 171
S. griseoplanus Antibiotics Ann. 1956-1957, 730 Alazopeptin
S. grisinus Folia Biol. 4: 263, 1958 Grisin
S. griseus var. farino- Bacteriol. Proc. p. 18, 1954 Streptolin
SUS
S. griseus var. spiralis Antibiotics Ann. 1959-1960, 194. Aspartocin
S. hepaticus Brit. Pat. 730,341, May 18, 1955
INCOMPLETELY DESCRIBED SPECIES OF ACTINOMYCETES 325
TABLE 27—Continued
Organism
Reference
Antibiotic
A. jucous
S. levoris
S. leydenematis
S. lilacinus
S. longisporus
S. luteochromogenes
S. luteolutescens
S. mediterranet
S. melanochromogenes
S.melanosporus (mel-
anosporofaciens )
S. natalensis
S. orchidaceus
S. paucisporogenes
S. phaeofaciens
S. phoenix
S. pleofaciens
S. plicatus
S. pluricolorescens
S. primycini
S. racemochromo-
genus
S. raffinosus
S. recifer
S. rutgersensis var.
casteranse
S. sakaiensis
S. salmonicida
S. sindenensis
S. subtropicus
S. toxicus
S. toyocaensis
S. vendargus
S. verticillus
S. vinaceus-drappus
A. violaceus-cristallo-
micint
S. viridifaciens
S. viridosporus
S. vulgaris
S. xcxanthochromogenes
S. zaomyceticus
Krassilnikov, 1955
Folia Biol. 4: 260, 1958; resembles S. griseus var.
candicidinus
Trans. Am. Microscop. Soc. 5: 376, 1953.
J. Antibiotics (Japan) 9B: 81, 1956
Folia Biol. 4: 263, 1958
J. Antibiotics (Japan) 6A: 188, 1953
Kurilowicz in Udientzev et al., p. 151, 1959
Antibioties Ann. 1959-1960, 262.
Kurilowiez in Udientzev et al., 1959
Giorn. Microbiol. 7: 207, 1959
Antibiotics Ann. 1957-1958, 878
Antibiotics & Chemotherapy 5: 204, 1955;
British Patent 768,007, February 138, 1957
Ann. Pharm. Franc. 16: 585, 1958
Japan. Med. J. 5: 327, 1952
Antibiotics & Chemotherapy 3: 788, 1953
Antibiotics Ann. 1954-1955, 806
Brit. Pat. Spec. 707,332, Apr. 14, 1954
J. Antibiotics (Japan) 9A: 75, 1956
Pharmazie 11: 304, 1956
J. Antibiotics (Japan) 9B: 170, 1956; LIB: 277,
1958.
Folia Biol. 4: 259, 1958
(Lima et al.) Morais, Lima and Maia, Syn. NV.
recifet Lima et al. (An. Soe. Biol. Pernambuco
15: 239, 1957)
Rev. Inv. Agr. Buenos Aires 8: 263, 1954
Ann. Rept. Takeda Research Lab. 14: 8, 1955
J. Bacteriol. 58: 659, 1949
Ann. Rept. Takeda Research Lab. 13: 41, 1954
Brit. Med. J. Nov. 12, 1955, Doklady Akad.
Nauk. SSSR 99: 827, 1954
Folia Biol. 4: 259, 1958
Japanese Pat. 32-3049, 1957
Australian Pat. 3985, Oet. 20, 1954
J. Antibiotics (Japan) 12A: 285, 1959
Brit. Pat. Spec. 708,686, May 5, 1954
Antibiotiki 2(5): 58, 1957
WLS? 2712 ope diulives 5s
770,065, Mar. 13, 1957
Brit. Pat. Spec. 712,547, July 28, 1954
Folia Biol. 4: 260, 1958
Bull. Agr. Chem. Soc. Japan 30: 469, 1956
J. Antibiotics (Japan) 7A: 134, 1954
1955; Brit. Pat. Spec.
Biomycin
Levorin
Cladomycin
Longisporin
Phthiomycin
Antitumor agent
Rifomycin
Antitumor agent
Melanosporin and
Elaiophylin
Pimaricin
Cycloserine
?
Phaeofacin
Rhodocidin
Pleomycin
Antibiotics C and D
Pluramyein
?
Racemomycin
Camphomycin
Monilin
Allomycin
Albomycin (grisein)
Necrotin
ml :
Toyocamycin
Oxytetracycline and
Vengicide
Pleomycin
Amicetin
Crystallomycin
Tetracycline and/or
chlortetracycline
Sistomycosin
Pneumocin
Xanthicin
Zaomycin
326
ar Cn OR, a ee Cy ee tee ees Teer Nee Ree ee ite aE heey ‘ beer Wi eh rye Pe) pt Tee APRS Cee, tee ore mn
, . : : : : QI c RS
BeeePeeee eee eeePeeePpPBeeeeeeepBepPapABeALBAAREABARARAARRARRER REE,
rs
»
‘
i
b b k
rs
nN
TABLE 28
. abikoensum var. spiralis
. acrimycint
. acrimycini var. globosus
. albidus var. invertens
. alborubidus
. albovinaceus
. atroolivaceus
. durentiogriseus
. aurint
. badius
. bicolor
. biverticillatus
. candidus var. alboroseus
. chromofuscus
. clnnabarinus
. coelicolor var. flavus
. coeruleofuscus
. coeruleorubidus
. coerulescens
. coerulescens var. longisporus
. cremeus
. cyanofuscatus
. daghestanicus
. flaveolus var. rectus
. flavotricini
. flavidovirens
. flavidovirens var. fuscus
. fradiae var. spiralis
. fumanus
. glaucescens
. glaucescens var. badius
. globisporus var. caucasicus
. globisporus var. flavofuscus
. gobitricini
. griseoloalbus
. griseoincarnatus
. griseomycini
. griseorubens
. griseoruber
. griseorubiginosus
. griseostramineus
. Wverini
. kurssanovii
. lateritius
. Litmocidini
. malachiticus
. mutabilis
. NUgreSCENs
. olivaceoviridis
THE ACTINOMYCETES, Vol. II
Incompletely described Streptomyces species of
Gause et al. (1957)
. clnnamonensis var. proteolyticus
. coelicolor var. achrous
. griseorubiginosus var. spiralis
TABLE 28—Continued
1. prunicolor
1. roseofulvus
1. roseolilacinus
{. roseolus
{. roseoviolaceus
1. roseoviridis
A. rubiginosohelvolus
1. rubiginosus
1. syringini
A. toxytricini
4. umbrinus
1. variabilis
1. variabilis var. roseolus
1. venezuelae var. spiralis
4. violaceorectus
1. violaceus var. rubescens
{. violascens
1. viridoviolaceus
tions of “new” organisms to emphasize the
ability of the cultures to produce melanin
pigments, an important and characteristic
property for distinguishing and identifying
species of Streptomyces, as brought out in
the various editions of Bergey’s Manual. In
spite of the fact that these descriptions of
“new” organisms had been undertaken for
the purpose of identifying antibiotic-pro-
ducing cultures, no attempt was made to
list any antibiotic for the various “new”
species; all one finds are such expressions
as: “strong repression,” ‘‘weak activity,”
‘no activity.” Even the antibiotic ‘“‘al-
bomycin” that has been greatly publicized
by the senior author is not listed in this
treatise; neither is the species A. subtropicus,
which produces this antibiotic, described.
Very little effort has been made in creating
these species to compare the newly isolated
cultures with those previously described and
well known in the literature. Many of the
new descriptions remind one of known organ-
isms.
For these reasons, this collection of ‘new’
species is listed here (Table 28) as incom-
pletely described, or at least as requiring
further information for proper identification.
Appendices
Two appendices are given here, each of
which is of special significance.
I. Since the colors of the substrate growth,
the aerial mycelium, and the spores, as well
as the pigment dissolved in the medium, are
highly important in classifying actinomycetes,
a well-recognized and universally available
standard must be used for color evaluation.
Unfortunately, some of the best standard
charts and volumes are not generally ac-
cessible. Hence, a simplified system, readily
understood, must be used. Such a system has
iets
presented here, with certain modifications
and in terms of English
as Appendix I. Several color charts are
available for this purpose. The following
standards may be consulted: Ridgeway
(1912), Sacchardo (1912), Maerz and Paul
(1930), and Munsell.
II. A knowledge of the chemical com-
position of the media used for the growth of
actinomycetes is essential for characteri-
zation of species and varieties. Suitable
media also are essential for good sporula-
tion and for the production of important
biochemical products, notably antibiotics
and vitamins. Numerous media have been
proposed for the growth of actinomycetes,
to serve purpose or another. To list
them all here is hardly necessary. Only a
few have been selected. These include syn-
thetic, artificial organic, and complex natural
media.
For examination of the
culture, it is desirable to make stained
preparations. Some of the general principles
underlying light microscopic and_ electron-
microscopic preparations have
cussed in Chapter 2 of Volume I.
been proposed by Lindenbein (1952).
equivalents,
one
microscopic
been dis-
Appendix I
Color Designations for Describing
Actinomycetes (Lindenbein)
1. White: (a)
white (candidus), (c) silver-white
(argenteus), (d) milk-white (lacteus), (e)
chalk-white (cretaceus), (f) gray-white (far-
inaceus); also cream, egg-shell, and ivory.
2. Violet: (a) bluish-violet
(b) reddish-violet (/2laceus), also lavender,
mauve, purple.
3. Blue: (a)
blue
snow-white (nzveus), (b)
elossy
(violaceus),
dark
(cyaneus), (Cc)
(azureus), (d) gray-blue
vellowish-blue (lividus).
(caeruleus), (b)
sky blue
(caesius), (e)
blue
cornflower
(viridis), (b)
(smaragdinus), (ce) blue-
green (glaucus), (d) forest-green (prasinus),
(e) olive-green (olivaceus).
5. Yellow: (a) light yellow (flavus), (b)
deep (luteus), (ce) eitron-yellow
(citrinus), (d) golden yellow (aureus), (e)
sulfur-yellow (sulfureus), (f) white-yellow
(stramineus), (g) brownish-yellow (gilvus),
(h) egg-yolk yellow (vitellinus), (i) pale
yellow (luridus), (k) greenish-yellow (galbus).
4. Green: (a)
emerald
grass-green
green
yellow
6. Orange: (a) light orange (aurantiacus),
(b) dark reddish-orange (croceus).
7. Red: (a) dark red (ruber), (b) carmine-
red (puniceus), (c) scarlet red (coccineus),
(d) fire-red (agneus), (e) pale carmine (roseus),
(f) flesh-red incarnatus), (g)
purple-red (purpureus), (hb) cinnabar-red
(cinnabarinus), (1) lead-red (miniatus), (k)
brick-red (lateritius), (1)
guineus), (m) brownish-copper red (cupreus),
(n) hght (rutilus); also pink,
coral-pink, rose, and wine-colored (vinaceus).
§. Brown: (a) light
(carneus,
blood-red (san-
vellow-red
brown (brunneus),
328 THE ACTINOMYCETES, Vol. II
(b) dark brown (umbrinus) (c) chestnut
brown (badius), (d) reddish-brown (fuscus),
(e) yellow-rusty-brown (ferrugineus), (f)
greenish-brown (hepaticus), (g) cinmnamon-
brown (cinnamomeus); also beige, tan, and
ocher.
9. Gray: (a) greenish-gray (griseus), (b)
ash-gray (cinereus), (c) white-gray (7ncanus),
(d) brownish-gray (fumzgatus), (e) reddish-
gray (murinus), (f{) bluish-gray (plumbeus).
10. Black: (a) gray-black (niger), (b)
coal black (ater), (¢) brownish-pitch-black
(piceus), (d) greenish-jet-black (coracinus),
(e) blue-black (atramentarius).
Appendix II
Certain Important Media for the Study
of Actinomycetes
Ixrainsky (1914) and Waksman and Curtis
(1916) were the first to report on the sig-
nificance of simple synthetic media in the
study of the morphological and cultural
properties of actinomycetes. Conn (1921)
made a careful comparison of the growth of
75 cultures of actinomycetes on a large
number of media. He came to the conclu-
sion that ‘‘extreme variation in chromo-
genesis is possible with some of the cultures
studied, according to the composition of
the medium, and some cultures even vary
greatly when studied at different times on
the same medium. The appearance of one
of these organisms on any medium should
not be described until it has been cultivated
on several lots of this medium at different
times. No culture, moreover,
sidered nonchromogenic until it has been
studied on a great variety of different pro-
tein-free media.”’
‘an be con-
The following have been selected as rep-
resenting the more important media recom-
mended for the study of actinomycetes. All
constituents are reported in grams per liter.
1. Sucrose nitrate agar:
DUCKOsear cca ate a ameter: 30.0 gm
INGINO eect orion aes pee 2.0 gm
| Cal nl) 1G Ae ne ie, wee Cree au SF 1.0 gm
WMeSO.e(HsO:s. 22.206: 0.5 gm
156 Ct seer eerie ce Wen ee he 0.5 gm
Bes Olas ote airs wens 0.01 gm
INGE wi) ope ae lee yo Miachae 15.0 gm
Distilled water........ 1000.0 ml
ple S060 13
This medium is frequently also known as
“Czapek’s agar,”’? or as ‘‘Czapek’s solution
agar.”’ Glycerol or glucose may besubstituted
for sucrose, giving glycerol-, or glucose-ni-
trate agar. Ammonium chloride may be sub-
stituted for NaNO; , giving sucrose-ammo-
nium agar.
2. Glucose-asparagine agar:
GIICOSES * sia. bb tna sauenaltatts 10.0 gm
ASpara eI. ...4 bocca. yen eet 0.5 gm
KEP On eks acer 5 el eee 0.5 gm
BP OT eS soitce, teks 6 eaten oe 15.0 gm
Distilled water........... 1000.0 ml
pH 6.8
Meat extract (2.0 gm) may be added to this
medium. Tap water may be used.
3. Glycerol-asparagine agar:
Gibyccnol dex. as ae eee 10.0 gm
ASPATAGINE. 2.6... osetia 1.0 gm
ESP Oreaco tamara oh, oo eae 1.0 gm
NaN) eee ea pee ae ee Ree ar Pfs 20.0 gm
Nea 1000.0 ral
Adjust to pH 7.0 with NaOH.
4. Glycerol-asparaginate agar I:
Glycerol. sche naneaecte 35.0 gm
Ammonium lactate........ 6.5 gm
Sodium asparaginate....... 3 ovom
IE RO) nForce 2.5 gm
INO eee 8 cere ten 2 epee = 5.0 gm
1, Gy erent irre tte are are a 0.1 gm
NaS Ogee. elise eciw See ae 0.3 gm
INIA Was epee eit. eee ene 20.0 gm
1000.0 ml
This medium is often spoken of as Ushin-
Distilled water. o.4.2.4-...<.
sky’s, especially when used as a solution,
without agar.
5. Glycerol-asparaginate agar IT:
GiivGeroly naiee cake eae 10.0 gm
Sodium asparaginate...... 1.0 gm
APPENDICES
K,HPO, arr Ree cemoe 8 = eas erlots Sao: 1.0 gm
NCU Rene tee ea eo 15.0 gm
Distilled water...... 1000.0 ml
This medium is often known as Conn’s.
6. Glycerol-glycine agar:
IVeCerOlepe re So oe shee 20.0 gm
(Glycine Anwar 64 2s 2.0 gm
JKGISN EQS Gt ees eee mee 1.0 gm
IEG) SR a ae ar re 2.0 gm
NTE SOR MESO. fossa 0.5 gm
RES Oz ey tices G5 so x sees O.1 gm
CAC Orme sirna css kbs wees 0.2 em
PMC See ce fo tar dks! SPUN ME 18.0 gm
Distilled water........... 1000.0 ml
Adjust to pH 7.2 with NaOH
This medium is often known as Plotho’s.
?. Glycerol-calcium malate agar:
CAINICCEO We lice IS a hitae Se 10.0 gm
Caleimm malate... ..nc 22: 10.0 gm
INGE © likay 4., Bch acts sas ates: 0.5 gm
ltl al 2G e Beeeeielae. gehen ae ey 0.5 gm
UNG ete a Surses to Acta. Ray chp 15.0 gm
Distilled water:......:.<. 1000.0: ml
Glucose or mannitol may be used (20 gm)
to replace the glycerol. Calcium citrate may
be used to replace the malate.
§. Glucose-ammonium salt agar:
CICOSeie eet ees sae 10.0 gm
GN) SEURORS... s..4-ahea5 4.0 em
INTGK Oe ey ae 5.0 gm
1] 6 0) Bk 0) i ae are Sa 2.0 gm
MGSO CHO a5 3c oc cerns 1.0 gm
CAG ete hs, was eS os oe 0.4 gm
Hes@ie dio Orin: f: a. 5 0.02 gm
WimsOue fiblsO. es. Joo 0.01 gm
INO REE SIT I's cha 15.0 gm
1000.0 ml
9. Glycerol-ammonium salt agar:
Distilled water...........
GIViGerOlin dane = aataue fea 10.0 gm
Il a ly G) ae las Sinan nage ae ng 1.0 gm
15S He One 1.0 gm
0.5 gm
>
Za
ne
2
a
>.
4
=
S
a |
5 gm
COO eye e arent hee 1.0 gm
ENO Trae Dost) ge eNiy sn = BA 15.0 gm
329
Distilled waiter.) sa. 7 0002 0emil
pH 7.0
The ammonium salt may be replaced by
KKNO;. This medium was proposed by
Masumoto (1952).
10. Glycerol-urea agar:
Glycerol 724. 4: arte ee 15.0 gm
Wrens 4. lato ia see ee 2.0 gm
Ke O 2. yeti ea 0.5 gm
INTE SOA Fs ede ans ue tomer 0.5 gm
NS Cte is lo Ohh oo 0.5 gm
HES Oe Misriens has mee 0.1 gm
ENE) IA Oe eee ERE a Ona y 15.0 gm
Distilled water: «52.0. se.
11. Glucose-tyrosine agar:
GrliGoses 7 4.9 oan Her ee 10.0 gm
‘Eyrosiie: Oi Saee oe eee 1.0 gm
CIES OM fakin serine alee 0.5 gm
GELB Ose xk xe oitangt Abe 0.5 gm
INGA T Rss be SPs cet ae Be goed Be 15.0 gm
Distilled water
teaction made neutral with NaOH.
12. Glucose-peptone agar:
Glucose, crude 40 gm
10 gm
PEplOne cso passe eee
Agar. powdereds =. 4 24208
Distilled waters =..5: sce
pH 5.6
Dissolve glucose and peptone in 500 ml dis-
tilled water. Dissolve agar, by heating at
100°C, in another 500 ml water. Mix the
two solutions, cool, add 10 gm egg-albumin
dissolved in 50 ml water. Shake, steam for
30 min, allow clot to settle, filter, distribute
in tubes, and autoclave at 115°C for 10 min.
This medium, also known as Sabouraud’s
agar, is fairly acid.
20 gm
1000.0 ml
13. Tyrosine-casein-nitrate agar:
AVPOSING aid te Sete S ae Oo
Sodium caseinate..........
Sodium nitrates. see.
LCE Ae ae a, Tee ae
1.0 gm
25.0 gm
10.0 gm
15.0 gm
Tapmweiler..2:2.. 55... 252 8 LOUORO ml
This medium has been recommended (Men-
zies and Dade, 1959) for the isolation of
Streptomyces scabies from potato scab lesions
500 THE ACTINOMYCETHS, Vol. II
or from soil. It inhibits spreading bacteria
and favors the production of a dark brown
pigment closely encircling colonies of the
pathogen. Since almost all pathogenic iso-
lates of S. scabies produce a dark pigment,
the selection of the probable pathogen from
dilution plates containing other actinomy-
cetes is favored.
14. Peptone-beef extract or nutrient agar:
PEPDOMNG 2 tbs. jee ome als ether 5.0 gm
BGEISEXUEA CU oi incon = 5.0 gm
NBIC] ees eh oo cath noel’ oa 5.0 gm
J3\(037) ie AA ee a 15 to 20 gm
Distilled water....... 1000.0 ml
pH 7.2 to 7.4
Irrequently 10 gm of peptone is used. Glu-
cose (10 gm) or glycerol (15 gm) may be
added to give glucose- or glycerol-peptone
agar. Tap water is often used in place of
distilled water. In a liquid state without
agar, these media are designated as nu-
trient broth, glucose-peptone broth, or glyc-
erol-peptone broth, respectively.
15. Peptone-beef-salt agar:
Gincose, oa Maan O22 oe eae 10.0 gm
BEDtONG: Sou ae. Sa Anes 5.0 gm
Beehextract. a. i.s4% eo... 5.0 gm
ERE Oi. geo pede Aus te fe: 1.0 gm
MesOue (sO ics ola tGeeee. 0.5 gm
SSC Ee ee oe a: eer Meee ere 0.5 gm
NCAT Meas eG is ls PE 15.0 gm
1000.0 ml
The organic nutrients may be reduced to
Distilled water.......
one half or even one tenth of above concen-
tration, in order to favor production of aerial
mycelium in some cultures of actinomycetes.
16. Glycerol-peptone-beef agar:
(li Genoa 4) eae da attest) ee 20.0 gm
PeDtONG Yea Rae a Hae d-8 5.0 gm
Beef extract. ........4.04. 3.0 ¢m
S\N eee te ee ee 15.0 gm
Distilled water............ 1000.0 ml
pH 7.0
17. Tryptone-yeast agar:
CEP DUONG peak ths ac aha ten acs 1.0 gm
VGast:extlacbs cule cas aa Ys 1.0 gm
5 gm
NTO ee roe gdp) Sys 5 MSE 17.0 gm
Tap water... ..... 24's... 00020) mal
18. Glucose-yeast-ammonium agar:
GIUCOSE ss 673 tors acitaay ote 10.0 gm
WeAast Extract. ocd o.ss6 ees 1.0 gm
GNP is ERP Oe e7) vonsn ce 1.0 gm
| 50) MORE een ae en ee ee 1.0 gm
MigSOi: (20 os c aee ese ees 0.2 gm
PREIS ES Ne enter 5 veal a on Pee 15.0 gm
Wistilled water.....:.:.... 1000°O<ml
pH 7.0
19. Gelatin media:
Crelaitiny :2ey csr udereee ee 160-200 gm
Tap water... <.06 +. 2425 “OOO 0mm!
Adjust to pH 7.4
Sterilize 30 minutes at 110°C
This medium may be supplemented with
glucose (20.0 gm) and peptone (5.0 gm) to
give glucose-peptone gelatin.
20. Gelatin agar medium:
PEPtONC e220 04s Manes Beas 5.0 gm
eel extrach. 2 .5..%4 450 e8 3.0 gm
Gelatin. < oc 24 2oet ees 4.0 gm
INGA os Saath aie Meee 15.0 gm
Distilled waiter:.u.45 afs oe. 1000.0 ml
pH 7.0
21. Starch agar:
Soluble starch. e..wese . 10.0 gm
INAIN OY oa bets hice erases 1.0 gm
KGEUP Os. 22 es nacre 0.3 gm
INVa Olah 1c tote cee eee 0.5 gm
INN Oy oi ee ets 2 is Se 1.0 gm
INGOT. Aad tele & oP aiecg: Gacy Ce 15.0 gm
Distilled. water: .......... 4. 1000.0 onal
CaCO; (3 gm) and MeSO,-7H2O (1 gm) may
replace the MgCOs;. (NH4)2SO. (2 gm) or
asparagine (0.05 gm) may be used to replace
the nitrate.
22. Starch-casein agar:
Soluble starch. .2...2hs244 10.0 gm
Casein (dissolved in NaOH) 1.0 em
eat Ons 5 as. Bact soe 0.5 gm
INGO S 2. 3. 56 w hue Meee eae 15.0 gm
Water. .<.t28 ste eaten 1000.0 ml
DEL (0 t6.:720
APPENDICES S51
23. Starch-peptone-beef-agar:
Starchiate sss ome ere... 10.0 gm
ReDuOner et toca ee es 5.0 gm
Mieatrextractes: 550s... 3. 3.0 gm
TAN RS isc ce hae ORE cae ee el rea 15.0 gm
Distilledwater........... 1000.0 ml
pH 7.0
24. Glycerol-starch-glutamate agar:
Givicerolmerc ak Mit ss: 10.0 gm
SEDIAG Ose ity Oo ele ek eee 10.0 gm
Sodium glutamate........ 1.0 gm
INAINO errors Oh els Seer. e 0.5 gm
FNOMMEn ge heres ols flee Sts Re 0.25 gm
\VANEE N00 vI@ eS 2 See ee 0.01 gm
Ibu Ose Sere 0.25 gm
Wc Oi ta elk 8 ee ae 0.25 gm
Kes Ona. «hee eke 0.01 gm
SWART es aed te Re as 15.0 em
Distilled water... ......=.. 1000-04
pili 0tor/s2
Shinobu (1958b) considered this one of the
best synthetic media for growth of actino-
mycetes, especially for production of aerial
mycelium.
25. Egg-albumin agar:
GlmCOseee sis aston o oek 10.0 gm
Reevalloumimin os 28: 230.4. 0.15 gm
GO eretteeee 0s ee 0.5 gm
Ne Oe Tain One, 2 otek 0.2 em
Bests @))seemenas cay fee Trace
EIS RG ON RM Be 15.0 gm
Distilled water......2...2. 1000:0mml
Ege albumin is first dissolved in water and
made neutral to phenolphthalein with N/10
NaOH.
26. Potato-glucose agar:
Peeled potatoes........... 200.0 gm
GIMIGOSES ete ae. eee 8S 20.0 gm
CAC Ons iee pearl: 0.2 gm
MeSOpaTHsOn aos cee: 0.2 gm
INOUE e Me Vs es ee eed oes 15.0 gm
Tap or distilled water...... 1000.0. ml
pH 6.8 to 7.2
27. Potato-peptone-glycerol agar:
OU aL On ee eee: Te ee aaa 100.0 gm
RepuOnesn ees 2.25 Sees e 2.0 gm
Glycetoles ys. 2. 5.0 gm
MicS O74: 3° penne eee 0.5 gm
KCHR Oe eee 0.5 gm
IN'as Ce Ae ae eee nag 0.5 gm
HeSOue oe :.4 Seer or eee 0.01 gm
AGO nate ae eters 15.0 gm
Mapswateres sss sone oe. LOUOZOmmn!
The potatoes are cut into small cubes to
which 100 ml of water is added and the
whole steamed for three quarters of an hour.
The extract is strained through fine muslin
without squeezing the pulp. The other nu-
trients are dissolved in 500 ml of water which
is then added to the potato extract, and the
whole steamed for three quarters of an hour.
The mixture is then made up to bulk, stand-
ardized and filtered, after which the agar is
added.
28. Glucose-yeast extract-beel-peptone agar:
GIMCOSeR Af. Aas tac inns: Bee 10.0 gm
10.0 gm
Beek extrachag. ows s--.4eac 4.0 gm
PEDUONER aah kh) eer ance: 4.0 em
NAC en een race eee eae 2.5) om
Distilled waters. 1000.0 ml
This medium is often known as Emerson’s.
Various modifications of this medium are
used. The concentration of the first two
constituents may be reduced to 1.0 gm per
liter; the NaCl may be left out.
29. Glucose-yeast extract agar:
Glucose 10.0 gm
10.0 gm
15.0 gm
1000.0 ml
AAD WabeD.) cam eek ie
pH 6.8
This medium
K.HPO,.
may also contain 0.5 gm
30. Glucose-casein digest-yeast-beef agar:
Glucose .s ents oe ele,
10.0 gm
1.0 gm
1.0 gm
2.0 gm
15.0 gm
1000.0 ml
Weast-extractaa «lees 42 os
Beetextract. - 4 ee
NeZ-AmInes A. -...8.. cee.
Distilledwaters.2. 22)...
pi 7-3
we)
bo
This medium is usually known as Bennett’s
agar.
31. Glucose-yeast-malt agar:
Grlui@OSe casi. Make Mele er eer: 4.0 gm
Weastiextracten ey 0s: 4.0 gm
Mita Re Xtraete icy ee en 10.0 gm
NOTE eee setla srsbeucena yi Oe cal TRU 20.0 gm
Distullediwaters:.cme.2.5. o- lO00Z0uml
pH 7.3
32. Dextrin-casein digest agar:
Dextre ea separ ees 7s 3 10.0 gm
Weast extractrnws f..sa.nck 1.0 gm
Beeiveximact.>. ks2e derkle 1.0 gm
Casein digest (N-Z-Amine A). 2.0 gm
CoG aie Oi. os eee 0.02 gm
20.0 gm
Distilediwater?.....:¢. 2225) 10005 0eml
Adjust to pH 7.3
This medium is often known as Hickey and
Tresner’s agar.
34. Oatmeal agar:
Rolkedvoaits 2). ah sina gee 20 to 65 gm
ita waber. = x. a8 spat on 100020Gml
Cook to thin gruel in double boiler, filter
through several layers of cheesecloth, and
make up to a liter while still hot. Add 18 to
20 gm agar. Adjust to pH 7.2 with NaOH.
34. Tomato paste-oatmeal agar:
20.0 gm
20.0 gm
500.0. ml
Add these two ingredients to the 500 ml of
Heinz baby oatmeal food... .
Tomato Paste ss 14 26.25 4 oa
Pap Wathen bets: St ts ht
boiling tap water. Do not adjust pH.
DiCarlo aera ees LosO vom
ABAD Wallet. .2 cucu fete 500.0. ml
Melt by steaming at 100°C for 15 to 20 min.
Do not adjust pH. Mix the two solutions,
steam at 100°C for 10 min, dispense, and
sterilize for 15 min at 121°C.
35. Soil extract agar:
IBGChOxtlaiGus. aren te sees
3.0 gm
PepuGne'sso5 24 ts tan ae Ses 5.0 gm
INOTS «oe eda een neh as ee 15.0 gm
SOMNERCEACh. 62g bs ae was 1000.0. ml
pH: 7.0
THE ACTINOMYCETES, Vol. II
The soil extract is prepared by treating | kg
garden soil with 2.5 liters of tap water for 1
hour in autoclave at 15 lb pressure. Filter
hot. Add tale for clarification if necessary.
36. Carbon nutrition medium:
Carbon source......... 10.0 gm
CNIPD) 0 One eee ee eas 2.64 gm
IPO ae eee 2.38 gm
KOEI RO.) aa: ee: 5.65 gm
MigS Occ (Hb Oe see 1.00 gm
CusOmoko On. 2 eae 0.0064 gm
RESO (Hs On ee cae 0.001 gm
Mn@ls=4i Oo a8 See 0.0079 gm
TmsSOie(H>oOs 2k Aer 0.0015 gm
ANON Chae ae eel OP eet 15.0 gm
Distilled water... .. 7 100020mnl
Some carbon compounds may have to be
sterilized separately either by filtration or
by heating in aqueous solution. This me-
dium was used by Pridham and Gottlieb
(1948), and has found general application
in the study of utilization by actinomycetes
of different carbon sources (Fig. 66).
37. Medium for nitrate reduction:
EDLONE: de oe ee eee eae 5.0 gm
Mieativextractecc ft. ase 3.0 gm
TRIN GOs. e258 2 ae, ae ee 1.0 gm
Distilled water: 02° 2 2: . 000020
pH 7.0
38. Medium for hydrogen sulfide production:
Peptone-iron agar (Difco)... 36.0 gm
Yeast extracte 2+: oe ee 1.0 gm
Distilled water 27% «026i. 1000.0 ml
39. Cellulose medium:
Filter paper saturated with synthetic
solution, free from other carbon sources.
40. Potato plug:
Plugs of potatoes of desirable size and
shape; distilled water added and steri-
lized.
The importance of the variety and health of
potato has been discussed by Grein and
Kuster (1955).
41. Milk or litmus milk:
Skim milk powder is used. Litmus or
brom-cresol may be added.
APPENDICES
Control (No Carbon) 1% Glucose 1% Sucrose 0.15% NaOAec
c f (
= an = fay
Streptomyces Sp. 10 Days at 28°C
FIGURE 66. Growth responses of streptomycetes, on chemically defined medium, to various
carbon sources (Courtesy of T. G. Pridham
42. Melanin formation medium: Use 4 to 5 ml portions in test tubes. Incubate
Yeast extract . l.0gm_ at 27°C and read after 1 to 2 days and after
L-Tyrosine. . 1.0gm 4 days.
NaCl... 8.5e@m 43. Other media:
Agar. >... 16.0 em A variety of other media are frequently
Tap water... 1000.0 ml used, often depending upon the nature of the
304 THE ACTINOMYCETES, Vol. II
organism and the problem. This is true par-
ticularly of the pathogenic forms, of organ-
isms not growing readily upon ordinary
media, and of those used for special purposes,
such as antibiotic production. Among the
complex media may be listed certain egg
media (Dorset’s), blood agar, blood serum,
‘carrot plug, and others (Waksman, 1919;
Levine and Schoenlein, 1930; Pridham et al.,
1956).
References”
Rules of
Gen.
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Aso, K., Aral T., YANAGISAWA, F., AND NaKa-
gima, M. Studies on the distribution of actino-
mycetes and their antagonistic strains in Japan-
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Aumauist, E. Untersuchungen iiber einige Bak-
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ANDERSON, L. E., Exuruicu, J., Sun, 8. H., ann
BURKHOLDER, P. R. Strains of Streptomyces,
the sources of azaserine, elaiomycin, griseo-
viridin, and viridogrisein. Antibiotics & Chemo-
therapy 6: 100-115, 1956.
Aoki, M. Agglutinatorische Untersuchung von
Aktinomyzeten. Z. Immunititsforsch. 86: 518—
524; 87: 196-199, 200-201; 88: 60-62, 1935-1936.
Avery, R. J. AND BuaNnk, F. On the chemical com-
position of the cell walls of the Actinomycetales
and its relation to their systematic position.
Can. J. Microbiol. 1: 140-148, 1954.
Backus, E. J., Duacar, B. M., anp CAMPBELL,
T. H. Variation in Streptomyces aureofaciens.
Ann. N. Y. Acad. Sci. 60: 86-102, 1954.
Baupaccr, E. La denomination ‘‘Actinomyces
bovis”’ Harz doit etre supprimée comme ‘‘nomen
dubium.’’ Bull. Sez. Ital. Soe. int. Microbiol.
8: 99-101, 102-105, 1936; 9: 138-146, 1937.
Baupaccr, E. Contributo alla sistematica degli
Attinomiceti V-VIII. Atti Ist. botan. Pavia
(4) 11: 191-231, 1939.
Baupaccr, E. Contributo alla sistematica degli
Attinomiceti. X-XVI Actinomyces madurae;
Proactinomyces ruber; Proactinomyces pseu-
domadurae; Proactinomyces polychromogenus;
Actinomyces violaceus; Actinomyces caeruleus;
con un elenco alfabetico delle specie e delle
varieta finora studiate. Atti. Ist. botan. Pavia
(5) 3: 1389-193, 1944.
Baupacctr, E. Criteria for the improvement of the
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Congr. Microbiol., Stockholm, 195s.
*Most of the references to the descriptions of
the various organisms listed in the text are at-
tached to the individual descriptions. Some of
the references listed here have also been given in
Volume I. For the sake of simplicity, they are
repeated here.
335
Baupaccit, E. Development in the classification
of actinomycetes. Giorn. Microbiol. 6: 10-27,
1958.
Baupaccr, E. anp Comascut, G. F. Contributo
alla sistematica degli attinomiceti: XVIII Ac-
tinomyces griseus (Krainsky) Waksman. Myco-
pathol. Mycol. Appl. 7: 278-281, 1956.
Baupacci, E., Comascui, G. F., Scortr, T., anp
Spauua, C. General criteria for the systematics
of genera and species of actinomyces (Strepto-
myces) and Micromonospora. VIth Intern.
Congr. Microbiol., Symp. Actinomycetales,
Rome, 1953, p. 20-39.
Baupaccl, E., Giuarpi, E., anp Amici, A. M.
Il ciclo di vita degli attinomiceti osservato al
microscopio elettronico. Giorn. Microbiol. 1:
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INDEX OF ORGANISMS
Boldface numerals indicate description of organisms.
Actinobacillus, 198 annulatus, 174
ligniersi, 320, 322 aquatilis, 316
paulotrophus, 35 arborescens, 316
Actinobacteriuwm, 20, 297 aromaticus, 316
abscessus, 20, 322 asteroides, 35
cellulitis, 15, 20, 322 asteroides var. serratus, 316
tsraelit, 17, 20 atrodiastaticus, 113
liquefaciens, 20 atroolivaceus, 123, 217, 326
meyeri, 20, 322 atypica pseudoturberculosa, 317
propionici, 2 aurantiacus, 90
Actinococcus, 198 aurantiogriseus, 271, 326
Actinomyces, vil, 1, 2, 4, 5, 12-20, 77, 95,496, 297, aureoverticillatus, 292
315, 316, 322 aureus, 87, oe
abikoensum var. spiralis, 166, 326 aurint, 237, ¢
acidophilus, 122 avadi, 317
acidoresistans, 316 baarnensis, 287, 317
acrimycint, 123, 229, 326 bacillaris, 218
acrimycint var. globosus, 123, 229, 326 badius, 287, 326
actinoides, 316 bahiensis, 317
actinomorphus, 316 baudetiz, 12, 13, 15
aerugineus, 316 beddardi, 122
africanus, 34 bellisari, 317
agrestis, 316 berardinist, 317
albidoflava, 316 berestneffi, 317
albidoflavus, 14, 120, 122, 123, 168, 197 bicolor, 131, 288, 317, 326
albidofuscus, 316 biverticillatus, 272, 326
albido-fuscus, 316 bobili, 87, 89
albidus var. invertens, 169, 326 nee ae res ole
alboatrus, 87, 316 bostroemi, 317
alboflavus, 87, 120 bovis, 12, 13-15, 17, 20, 29, 78, 278
albopurpureus, 316 bovis albus, 317
alborubidus, 122, 326 bovis farcinicus, 317
albosporeus, 86 bovis luteoroseus, 317
albovinaceus, 326 bovis sulfureus, 317
albus, 6, 71, 87, 90, 118, 119, 120, 264, 320 bovis var. nigerianus, 317
albus-acidus, 316 brasiliensis, 285
albus asporogenes, 316 bronchialis, 317
albus chlamydosporus, 172, 316 bronchiticus, 317
albus var. acidus, 316 brumpti, 317
albus var. alfa, 316 brunt, 317
albus var. fungatus, 172 buccalis, 317
albus vulgaris, 172, 316 caeruleus, 91
allenbachi, 316 Californicus, 87
almquisti, 119, 122, 316 cameli, 317
alni, 197 caminiti, 317
americanus, 316 candidus, 91, 122, 223
anaerobicus, 316 candidus var. eke 122, 188, 326
anaerobies, 316 canis, 20, 317
347
348 INDEX OF
canis familiarts, 317
caprae, 38
carneda, 317
crnews, 60
carougeaut, 317
catarrhalis, 3
cati, 317
caucasicus, 218
caviae, 317
cellulitis, 12, 15,8
cellulosae, 89, 192
cerebriformis, 317
cereus, 317
chalmersi, 317
christophersoni, 317
chromofuscus, 145, 326
chromogenes, 90, 144, 202, 208, 209, 235, 253, 273,
317
chromogenus. 6, 71, 87, 118, 119, 122, 144
cinereo-niger, 317
cinereonigeraromaticus, 317
cinnabarinus, 131, 326
cinnamonensis var. proteolyticus, 196, 326
circulatus, 91, 324
citrea, 317
citreus, 87, 196
citrotremeus, 317
cloacae, 317
coccocidus, arn
coelicolor, 90, 282, 283
coelicolor var. pie 326
coelicolor var. flavus, 326
coeruleofuscus, 131, O88, 326
coeruleorubidus, 131, 288, 326
coerulescens, 131, 288, 326
coerulescens var. longisporus, 288, 326
coeruleus antibioticus, 324
colorata, 317
congolensis, 317
convolutus, 318
coremiales, 318
cremeus, 170, 326
cretaceus, 91, 318
cruoris, 318
crystallophagus, 318
cuniculr, 318
cyaneus, 90
cyanofuscatus, 197, 218, 326
cylindraceus, 318
cylindrosporus, 91
daghestanicus, 196, 326
dassonviller, 318
decussatus, 318
denitrificans, 318
dermatonomus, 318
diastaticus, 86
ORGANISMS
diastaticus var. ardesicicus, 201
diastaticus var. venezuelae, 201
diastato-chromogenus, 87, 89
discofoliatus, 12, 15-16
dispar, 318
donnae, 318
dori, 318
equptt, 318
elaeagni, 318
elephantis primigeniz, 318
enteritidis, 318
equi, 318
erysipeloides, 318
erythreus, 122
erythrochromogenus, 87, 89
exfoliatus, 87, 122
farcinicus, 122
farinosus, 91, 188
fasciculus, 188
ferrugineus, 318
flava, 318
flaveolus var. rectus, 326
flavidovirens, 170, 326
flavidovirens var. fuscus, 170, 326
flavotricini, 235, 326
flavus, 87,90, 170, 179, 192, 197, 210, 213, 252, 318
flocculus, 122
fluorescens, 218, 318, 324
foulertoni, 318
fradiae, 90, 138
fradiae var. spiralis, 324, 326
Fradi, 87
freeri, 318
fulvisstmus, 89
fumanus, 196, 326
fumosus, 91
fusca, 319
fuscus, 319
gabritscheoskt, 319
gartent, 319
gedanensis, 122, 319
gelaticus, 122
genesil, 319
gibsoni, 319
glaucescens, 131, 218, 326
glaucescens var. badius, 218, 326
glaucus, 90
globisporus, 90, 218, 226
globisporus albus, 218
globisporus circulatus, 218
globisporus diastaticus, 218
globisporus flaveolus, 218
globisporus griseus, 218
globisporus lactis, 218
globisporus scabies, 218
globisporus streptomycini, 143, 218, 226
INDEX OF ORGANISMS 349
globisporus var. caucasicus, 326
globisporus var. flavofuscus, 326
globisporus vulgaris, 218
globosus, 91
gobitricint, 286, 326
goensis, 319
gonadiformis, 319
gougeroti, 119, 122
gracilis, 90
graminearus, 319
graminis, 319
griseoflavus, 89
griseoincarnatus, 227, 326
griseoloalbus, 170, 326
griseomycint, 123, 148, 326
griseorubens, 123, 217, 326
griseoruber, 131, 272, 326
griseorubiginosus, 131, 266, 326
griseorubiginosus var. spiralis, 266, 326
griseostramineus, 210, 326
griseo-viridis, 319
griseus, 87, 89, 90, 123, 133, 1385, 143, 218, 286
griseus variabilis, 123, 319
griseus zonatus, 123, 287
gristnus, 218
gruberi, 319
gqueguent, 319
guerrat, 319
guignardi, 319
gypsotdes, 319
halotricus, 319
halstedit, 87, 90
heimi, 319
heimat, 122
hobnesi, 319
hofmanni, 319
hominis, 14, 819
hvidhansent, 12, 16-17
incanescens, 285, 319
indicus, 319
innominatus, 319
interproximalis, 20, 319
invulnerabilis, 319
israeli, 14, 16
israeli var. indo-sinensis, 17
israelit, 12, 13, 16, 17-18, 19, 20, 31, 78
iverini, 326
japonica, 319
japonicus, 319
jollyt, 319
jucous, 325
keratolyticus, 319
kimbert, 122
krainskiz, 209, 319
krauset, 319
kurssanoviz, 231, 326
lacertae, 319
lanfranchii, 319
lasseret, 319
lateritius, 285, 326
lavendulae, 87
leishmani, 319
lepromatis, 319
levoris, 197, 218
levyi, 319
lieskez, 122, 320
liguire, 320
lingualis, 320
Lipmanii, 87
liquefaciens, 320
listerz, 122
litmocidini, 326
londinensis, 320
longisporus, 90, 122, 172, 218, 320
longisporus-flavus, 90
longisporus-fungatus, 172
longisporus griseus, 172, 320
longisporus-ruber, 90, 271, 320
longissimus, 91, 212, 320
luridus, 212
luteolus, 320
luteo-roseus, 320
macrodipodidarum, 320
malachiticus, 130, 287, 326
malenconi, 122, 320
matruchoti, 320
melanocyclus, 90
melanogenes, 248, 320
melanoroseus, 320
melanosporeus, 243, 320
metchnikovt, 320
mexicanus, 37, 320
micetomae, 320
microflavus, 320
mihi, 320
mineaceus, 320
minimus, 320
minutissimus, 320
mirabilis, 122
mishagiensis, 320
mordoré, 321
mucosus, 320
multifidus, 320
muris, 49
muris-ratti, 320
musculorum, 320
mutabilis, 320, 326
myricae, 320
naeslundit, 12, 13, 18-19
necrophorus, 320
neddeni, 320
neschezadimenki, 320
300 INDEX OF ORGANISMS
nicoller, 320 rectus, 91, 209, 281, 303
niger, 90, 247, 320 rectus brunneus, 281
niger aromaticus, 247, 320 reticult, 87, 91, 122
nigrescens, 231, 326 reticulus, 272
nigricans, 320 reticulus-ruber, 91, 272
nigrificans, 90, 247, 320 ribeyrot, 321
nitrogenes, 320 rivieret, 321
nocardit, 320 rodellae, 321
nodosus, 320 rogersti, 321
nondiastaticus, 320 rosaceus, 321
non-fluorescens, 320 rosenbachi, 321
ochraceus, 60, 320 roseochromogenes, 268
ochroleucus, 60, 275, 320 roseodiastaticus, 201
odontolyticus, 12, 19-20 roseoflavus, 133
odoratus, 320 roseofulvus, 270, 326
odorifer, 320 roseolilacinus, 286, 326
oidiosporus, 91 roseolus, 286, 321, 326
oligocarbophilus, 320 roseoviolaceus, 285, 326
olivaceoviridis, 130, 210, 326 roseoviridis, 286, 326
olivaceus, 89 roseus, 87, 89, 268
orangico-niger, 321 ruber, 90, 182, 204, 271, 321
orangicus, 321 ruber sterilis, 54
panginensis, 321 rubidaureus, 321
parvus, 87 rubiginosohelvolus, 143, 326
pelogenes, 321 rubiginosus, 128, 217, 326
penicillotdes, 321 rubrocyanodiastaticus, 118
phaeochromogenus, 89 rubrocyanodiastaticus var. impiger, 201
phagocidus, 321 rubrocyanodiastaticus var. piger, 201
phenotolerans, 90, 321 Rutgersensis, 86
pijpert, 321 sabrazés, 321
pinoyt, 321 saharae, 321
plurichromogenus, 321 salmonicolor, 90, 239
pluricolor, 90, 283, 321 salvati, 321
pluricolor diffundens, 259, 321 sanfelicer, 321
polychromogenus, 321 sanguinis, 321
ponceti, 321 sanninii, 119, 122
pranicolor, 131 saprophyticus, 122, 322
pretorianus, 321 saprophyticus var. chromogenes, 322
protea, 321 sartoryt, 322
prunicolor, 145, 326 scabies, 90, 197, 277
pseudonecrophorus, 321 scabies var. anglica, 322
pseudotuberculosae, 321 sendatensis, 322
pseudotuberculosis, 321 septicus, 322
pseudotuberculosus, 321 serratus, 322
pulmonalis, 321 setonit, 90
puntonit, 321 setonit flavus, 275
pur peo-chromogenus, 87 silberschmidtt, 322
purpureus, 321 somaliensis, 122, 322
purpurogenus, 87, 321 sommert, 322
putori, 49 spinae, 322
putridogenes, 321 sptnosporus, 322
putrificus, 321 spttzi, 322
pyocyaneus, 321 splenicus, 322
pyogenes, 321 spumalis, 90, 322
radiatus, 321 streptomycint, 148, 218, 226
raffinosus, 218 subtropicus, 220, 326
INDEX OF ORGANISMS dol
sulfureus lacertae, 319
sulphureus, 14, 278
syringint, 286, 326
tarozzi1, 322
tenuis, 322
thermodiastaticus, 119
thermophilus, 119, 122, 300, 301
thermotolerans, 322
thibrerget, 322
thjottae, 322
thuilleri, 32
tossicus, 322
totschidlowskiz, 90, 322
toxicus, 218, 322
toxytricini, 235, 326
transvalensis, 322
tricolor, 198, 322
tyrosinaticus, 322
umbrinus, 200, 326
upcottit, 122
urethritidis, 322
urinarius, 322
valvulae, 322
valvularis, 322
valvulas desiiens bovis, 322
variabilis, 266, 326
vartabilis var. roseolus, 266, 326
venezuelae var. spiralis, 281, 326
Verne, 87
verrucosus, 322
verticillatus, 91
verticillatus viridans, 282
violacea, 285, 322
violaceochromogenes, 145
violaceorectus, 131, 285, 326
violaceus, 90, 251, 282, 283
violaceus-caesert, 86, 131, 251, 285
violaceus chromogenes, 285
violaceus cristallomicini, 325
violaceus-niger, 87, 90
violaceus-ruber, 87, 89, 282, 283
violaceus var. rubescens, 285, 326
violascens, 326
virgatus, 90
viridans, 90
viridis, 91, 120, 130, 136, 236, 287, 317
viridis sterilis, 287
virtdochromogenes, 90, 172, 306
virido-chromogenus, 87
virtdodiastaticus, 201
viridiflavus, 288
viridoviolaceus, 131, 285, 326
vulgaris, sae
waksmanii, 282, 283, 322
willmorez, 12
xanthostromus, 275, 322
Actinomycetaceae, vit, 2
Actinomycetales, vit, 1, 2, 75, 316
Actinomycetes, incompletely described, 315-326
Actinoplanaceae, vit, 310-314
Actinoplanes, vi1, 4, 5, 81, 84, 310-313
philippinensis, 311-313
Anaeromyces bronchitica, 317
Asteroides, 323
liskeyi, 323
pseudocarneus, 323
Bacillus, 322, 323
actinoides, 322
bifidus, 323
coelicolor, 140
mycoides corallinus, 23, 40
ramosus, 323
Bacterium, 322, 323
actinocladothrix, 14, 322
Brevistreptothrix, 323
israeli, 17
Chainia, 5
Cladothriz, 315, 323
actinomyces, 14, 323
asteroides, 35
bovis, 14
dichotoma, 119, 122
foersteri, 318
tnvulnerabilis, 122
liquefaciens, 122
odorifera, 119, 122
rubra, 129
Cohnistreptothriz, 20, 323
americana, 316, 323
israeli, 14, 17
silberschmidti, 318
Corynebacterium, 1
acnes, 13
israeli, 17
Discomyces, 315, 322, 323
asteroides, 323
bovis, 14, 17
decussatus, 318
israeli, 17
lingualis, 319
pleuriticus, 322
pleuriticus canis familiaris, 322
Escherichia coli, 137, 174
Eubacteriales, 1
Flavobacterium, 323
salmonicolor, 323
Fungus sterilis, 153
Jensenia, 5
Lactobacillus, 1
Leptothrix oculorum, 318
Microbacterium mesentericum, 24
302 INDEX OF ORGANISMS
Microbispora, 5, 84, 298-299 atlantica, 33, 36, 37
amethystogenes, 299 aurea, 317, 318
amethystogenes var. nonreducans, 299 autotrophica, 179
chromogenes, 299 babiensis, 239
diastatica, 299 berestneffi, 317
parva, 299 bicolor, 317
rosea, 298 bifida, 323
rosea var. nonnitritogenes, 299 blackwelliz, 28, 33, 37
Micrococcus pelletieri, 256 bovis, 14, 30, 323
Micromonospora, vit, 1, 2, 5, 84, 293-297, 300, 301, brasiliensis, 27, 29, 32, 37, 38, 58, 183
306, 309 brumpti, 239
bicolor, 294-295 bruni, 317
cabaelli, 294 buccalis, 317
chalcea, 293, 295, 301 calearea, 32, 38
coerulea, 294, 295 candida, 317
elongata, 294, 295-296 caprae, 27, 32, 35, 38, 39
fusca, 293, 294, 296, 301 carnea, 29, 317
gallica, 293, 296 carougeaut, 317
globosa, 293, 294, 296 caviae, 33, 39
monospora, 294 cellulans, 32, 39, 40
parva, 294, 296-297 chalmersi, 317
propionica, 294, 297 christophersoni, 317
vulgaris, 294, 300, 301, 309 chromogena, 35
Micropolyspora, 2 citrea, 32, 40
brevicatena, 2 citreus, 31
Mycobacterium, vit, 5, 21, 23-25, 67, 322, 323 coeliaca, 27, 32, 34, 40
agreste, 23 convoluta, 318
albuvialum, 323 corallina, 23, 27, 30, 32, 40, 41, 45, 50, 53-56
crystallophagum, 23 crateriformis, 36
erythropolis, 28 cruoris, 318
opacum, 23 crystallophaga, 50
paraffinicum, 322 cunicult, 57, 318, 323
rhodochrous, 27 cyanea, 132
salmonicolor, 23 cyaneus, 31
tuberculosis, 137, 175 cylindracea, 318
Mycococcus, 5 dassonvillet, 318
Nocardia, vi1, 1, 2, 4, 5, 23-60, 66, 73-75, 77, 78, 81, decussata, 318
83, 121, 129, 216, 236, 247, 285, 309 diastatica, 35
acidophilus, 324 dicksonit, 33, 41
actinotdes, 30 elaeagnit, 30
actinomorpha, 24, 30, 33, 34 enteritidis, 318
actinomyces, 14, 23, 323 eppingert, 35
africana, 38, 34, 60 equi, 318
agrestis, 54 erythropolis, 24, 25, 27, 30, 50, 328
alba, 33, 34-35 farcinica, 23, 27, 31, 32, 41, 42
alba lactica, 35 fastidiosa, 27, 32, 41, 42
albicans, 30, 32, 35 ferruginea, 23, 318
albida, 323 filiformis, 323
albosporea, 323 flava, 32, 42
albus, 30 flavescens, 24, 30, 31, 33, 43
alni. 30% 33,/ 30 flavus, 31
anaerobicus, 31 foerstert, 28
appendicis, 323 ford, 338, 43
aquosus, 25 formica, 33, 44
arborescens, 23 forstert, 318
asteroides, 25, 27-29, 31, 32, 35, 36, 47, 67, 272, foulertoni, 318
318, 021 freerz, 31
fructifera, 31, 33, 44
fusca, 319
gabritschewski, 31
gardneri, 28
gartent, 319
gedanensis, 30, 319
genesit, 256, 319
gibsonit, 33, 44, 45
globerula, 27, 32, 45
goensis, 319
grubert, 319
gypsoides, 29, 36
hoffmanni, 35
hortonensts, 38, 45
indica, 256
indica var. flava, 319
intracellularis, 32, 45, 46
israeli, 17, 31
tvorensis, 33, 46
krainskii, 323
krauset, 319
kurotishi, 33, 46, 47
lanfranchit, 319
lasserez, 319
letshmanit, 27, 29, 32, 35, 47
lignieresi, 31, 32 ;
lingualis, 319
liquefaciens, 320
listeri, 33, 47
londinensis, 320
lurida, 324
lutea, 27, 33, 48
macrodipodidara, 320
maculata, 24
madurae, 239
marina, 33, 37, 48
mesenterica, 24, 30, 32, 48, 49
mexicana, 29, 320
minima, 27, 30, 35, 40, 54-55, 323
minutissima, 320
muris, 31, 33, 49
myricae, 30
narasinoensis, 324
nicoller, 320
niger, dl
nigra, 33, 46, 49
nitrificans, 248
oligocarbophilus, 30
opaca, 23, 24, 27, 30, 32, 49-51
panginensis, 321
panjae, 33, 51
paraffinae, 24, 27, 30, 32, 51, 52
paulotropha, 35
petroleophila, 32, 52
phenotolerans, 35
pijpert, 321
INDEX OF ORGANISMS
pinoyt, 321
plurichromogena, 321
pluricolor, 321, 32:
polychromogena, 24
polychromogenes, 25, 27, 30-32,
ponceti, 321
pretoriana, 27, 37, 321
pseudocarneus, 36
pseudotuberculosis, 321
pulmonalis, 27, 32, 53
putoriae, 31
pyogenes, 31
ramosa, 323
rangoonensis, 33, 53, 54
rhodnit, 27, 33, 54
repens, 323
rogerst, 321
rosenbachi, 318
ruber, 25, 31
rubra, 26-28
rubropertincta, 27, 32, 42, 55
rugosa, 28, 32, 55
salivae, 60
salmonicolor, 23, 27, 30, 32, 55, 36, 2
sanfelicer, 321
saprophytica, 323
saturnea, 60
sebivorans, 32, 56, 57
sendaiensis, 31
serophila, 32, 57
silberschmidti, 322
somaliensis, 31
splenica, 322
sumatrae, 33, 57, 58
sylvodorifera, 35, 323
tenuts, 322
thibierget, 322
thiryei, 321
thuilliert, 322
transvalensis, 27, 32, 37, 58
turbata, 33, 58
uniformis, 33, 58, 59
upcottit, 33, 59
vaccinit, 32, 59, 60
valvulae, 322
variabilis, 31, 32, 60
viridis, 31, 32, 60
Oidium lactis, 140
6, 315, 316
alpha, 122
Oospora,
anaerobies, 316
asteroides, 35
bovis, 14
buccalis, 317
caprae, 38
chromogenes, 317
48, 50, 52, 53
8, 33, 48, 54, 55, 25:
3D
3
354
INDEX OF ORGANISMS
cretacea, 318
cylindracea, 318
decussata, 318
doriae, 122, 318
forsteri, 318
hominis, 229
micetomae, 320
mordoré, 321
rosella, 321
rubea, 321
spitzt, 322
tenaz, 322
Polysepta, 5, 323
Proactinomyces, 5, 24, 323
actinoides, 324
aquosus, 323
asteroides var. crateriformis, 323
asteroides var. decolor, 323
atlanticus, 36
citreus marinae, 48
cyaneus, 323
cyaneus antibioticus, 323
flavus, 37, 42, 48
freeri, 318
israeli, 17
minimus, 40
murts, 49
opacus, 50
pseudomadurae, 323
restrictus, 323
Pseudonocardia, 5, 302, 309
thermophila, 308, 309
Serr
atia, 323
corralina, 323
Sphaerotilus bovis, 14
Streptobacillus moniliformis, 49
Streptomyces, vi, 2, 4-6, 8-10, 21, 22, 24, 25, 84,
293, 300, 301, 302, 303, 306, 309, 311, 314, 315,
322, 333
Characterization of, 61-81
Deseription of, 84
Groups of, 82-114
albus, 92, 120
griseus, 92
lavendulae, 92
parvus, 92
reticuli, 92
viridochromogenes, 92
Identification of, 82-114
Sections of
biverticillus, 97
biverticillus-spira, 97
monoverticillus, 97
monoverticillus-spira, 97
rectus-flexibilis, 97
retinaculum-apertum, 97
sptra, 97
Series of, 95, 96, 115-151
albidoflavus, 95, 124
albosporeus, 96
albus, 95, 96, 111, 117-123, 211, 217
antibioticus, 95, 118, 124
aureus, 95, 96, 124
azureus, 131
bostroemi, 95, 113
caeruleus, 95, 113
cas-gri, 144
chromogenes, 96
chromogenus, 144, 147
chrysomallus, 96
cinereo-ruber, 96
cinereus, 111, 117, 123, 148
cinnamomeus, 96, 112, 117, 149
circulatus, 96
coerulescens, 96
diastaticus, 95, 113
erythrochromogenes, 112, 117, 149
flavoviridis, 95
flavus, 95, 111, 117, 123-129
flavus-parvus, 128
fradiae, 95, 96, 111, 117, 131-133
fuscus, 96, 138
griseoincarnatus, 95
griseocarneus, 96
griseus, 95, 96, 111, 112, 117, 133-143, 229, 244,
260, 279
helvolus, 96, 148, 144
hygroscopicus, 112, 117, 124, 143, 144
intermedius, 95
lavendulae, 95, 112, 117, 144, 145-149
lavendulae-roseus, 96, 147
madurae, 95, 129
melanosporeus, 129
nigrescens, 96
phaeochromogenes, 110
reticuli, 96, 112, 117, 144, 149-151
rimosus, 95
roseochromogenes, 95
roseoflavus, 95
roseoviolaceus, 96, 131
roseus, 95, 129
ruber, 96, 111, 117, 129
rubrireticuli, 96
scabies, 95, 112, 117, 144, 145
sulphureus, 95, 124
thermophilus, 112, 151, 152, 302-304
verticillatus, 96
violaceoruber, 111, 117, 130, 131
violaceus, 96, 97, 131
virgatus, 95
INDEX OF ORGANISMS 3099
viridis, 95, 111, 117, 130, 286
viridochromogenes, 112, 117, 149
Species of, 115-151, 152-164, 165-292
abikoensum, 163, 165, 166, 267
aburaviensis, 156, 166
achromogenes, 158, 166, 167, 173
acidomyceticus, 163, 167, 196, 235
acidophilus, 159, 167, 168
aerocolonigenes, 291
afghaniensis, 163, 168
africana, 29
africanus, 256
albicans, 324
albidoflavus, 157, 168, 169, 232, 235, 268
albidofuscus, 264, 324
albidus, 159, 169
albireticult, 105, 150, 151, 1638, 169
alboflavus, 124, 160, 169, 170, 232, 235, 254, 255,
265
albogriseolus, 94, 99, 105, 158, 170
alboniger, 163, 170, 171
albosporeus, 129, 133, 160, 171
alboviridis, 130, 157, 171, 172
albulus, 324
albus, 29, 68, 93, 94, 102, 105, 109, 111, 117-121,
123, 156, 169, 172, 174, 219, 254, 287
almquisti, 219
althioticus, 158, 172, 173
ambofaciens, 158, 173, 177
aminophilus, 324
annulatus, 156, 173, 174, 187
antibioticus, 66, 101, 102, 105, 108, 142, 161,
174, 186, 205, 279
antimycoticus, 160, 174, 175, 246
arabicus, 324
arenae, 156, 175
argenteolus, 156, 175, 176
armillatus, 125, 159, 176
aurantiacus, 158, 176, 177
aureofaciens, 71-73, 93, 94, 101, 102, 105, 124,
127, 128, 157, 173, 177, 216, 274
aureoverticillatus, 292
aureus, 124, 162, 177-179, 228, 250, 279
autotrophicus, 156, 179
bacillaris, 324
badius, 324
beddardii, 161, 179, 180
bellus, 162, 180
bikiniensis, 68, 105, 161, 174, 180, 181, 228
blastmyceticus, 161, 181, 182, 324
bobiliae, 98, 130, 149, 162, 176, 182, 194
bottropensis, 158, 182, 183, 204
brasiliensis, 160, 183
cacaot, 94, 159, 183, 184, 254, 264
caelestis, 156, 184, 218
caeruleus, 105, 181, 157, 184, 185
caespitosus, 163, 185
caiusiae, 161, 185, 186, 234
californicus, 85, 93, 94, 105, 140, 141, 160, 186-
187, 268
calvus, 123, 156, 187, 188
candidus, 160, 187, 188
canescus, 68, 158, 188-190, 197, 198
canus, 160, 190
carcinomyceticus, 324
carnosus, 162, 190, 275
casei, 302, 303
catenulae, 156, 190, 191
cavourensis, 161, 19]
celluloflavus, 129, 158, 191, 192
cellostaticus, 324
cellulosae, 123, 157, 192, 248
chartreusis, 68, 93, 149, 162, 184, 192, 193, 288
chattanoogensis, 324
chibaensis, 158, 193, 324
chrestomyceticus, 324
chromogenus, 144
chrysomallus, 93, 102, 129, 141, 142, 158, 193, 194.
chrysomallus var. fumigatus, 102, 194
cinereoruber, 130, 149, 162, 174, 194, 195
cinereoruber var. fructofermentans, 130, 194
cinnamomeus, 117, 149, 163, 195, 247
cinnamomeus f. azacoluta, 195
cinnamomeus f. cinnamomeus, 195
cinnamonensis, 68, 105, 148, 161, 195, 196, 243,
250, 324
circulatus, 147, 150, 163, 196
citreus, 124, 159, 196, 197
clavifer, 145, 157, 197, 275
coelicolor, 70, 76, 78, 93, 102, 105, 106, 131, 132,
140, 157, 169, 190, 197, 198, 212, 282-284
collinus, 163, 168, 198, 199
colombiensis, 324
coroniformis, 156, 199
craterifer, 93, 117, 128, 156, 197, 199
cyaneofuscatus, 131
cyaneus, 93, 131, 149, 163, 199
cyanoflavus, 131, 132, 157, 199, 200
cylindrosporus, 162, 200, 262
dassonvillez, 130
decaris, 212
diastaticus, 93, 158, 167, 200, 201, 202, 239
diastatochromogenes, 94, 161, 201, 202, 222
echimensis, 105, 151, 324
echinatus, 64, 99, 163, 202, 203
elasticus, 156, 203
endus, 144, 160, 203, 204
erythraeus, 66, 100, 105, 129, 158, 176, 204, 291
erythrochromogenes, 117, 129, 149, 162, 198,
204, 205, 225, 291
eurocidicus, 163, 205
eurythermus, 161, 174, 205
lr
oo
INDEX OF ORGANISMS
exfoliatus, 159, 205, 206
farinosus, 94
fasciculatus, 324
fasciculus, 94
felleus, 158, 206, 252
fervens, 149, 163, 206, 207
filamentosus, 159, 207
filipinensis, 162, 207, 208, 212
fimbriatus, 145, 163, 208
fimicarius, 124, 158, 166, 167, 206, 208
flaveolus, 68, 93, 99, 105, 124, 125, 142, 157, 203,
208, 209
flavochromogenes, 94, 161, 182, 209, 215, 222, 250
flavofungini, 324
flavogriseus, 93, 129, 158, 209, 212, 224
flavoreticuli, 163, 210
flavovirens, 105, 129, 142, 158, 210, 236
flavoviridis, 237
flavus, 93, 117, 124, 127, 129, 159, 198, 210, 211,
213, 252, 254
flocculus, 158, 211, 264
floridae, 141, 186
fluorescens, 324
fordiz, 124
fradiae, 66, 75, 94, 100, 105, 117, 132, 133, 134,
147. 154. 160, 209,210; 211, 212, 213; 238,
268, 284, 324
fragilis, 158, 212
fulvissimus, 99, 102, 159, 213, 276
fumosus, 160, 213
fungicidicus, 156, 213, 214
fuscus, 133, 163, 214
galbus, 102, 161, 214, 215
galbus var. achromogenes, 102, 215
galilaeus, 101, 162, 215
galtiert, 145, 161, 215
ganmyceticus, 324
ganmycicus, 324
gardnert, 156, 215, 216
garyphalus, 162, 216
gedanensis, 159, 216, 217
gelaticus, 159, 217
glaucus, 159, 184, 217, 218
globisporus, 70, 93, 156, 218, 254
globisporus streptomycini, 145
globisporus tundramycint, 324
globosus, 93, 162, 218, 219
gougeroti, 159, 219
gracilis, 162, 219
graminofaciens, 324
griseinus, 8, 114, 140, 160, 219, 220, 226
griseobrunneus, 161, 220
griseocarneus, 105, 150, 151, 163, 220, 221, 243
griseochromogenes, 161, 221, 222
griseoflavus, 99, 124, 127, 157, 176, 208, 222,
237, 249, 277
griseolus, 93, 102, 105, 156, 222, 223, 248, 252,
262
griseoluteus, 105, 107, 158, 223
griseoplanus, 159, 223, 224, 324
griseoruber, 94, 162, 224, 225
griseoviridis, 130, 162, 225
griseoviridus, 225
griseus, 8, 29, 66, 68, 74-76, 83, 98, 94, 99, 105,
114, 117, 128, 135-141, 148, 159, 169, 171, 179,
188, 192, 194, 220, 222, 225, 226, 236, 253,
256, 269, 275
griseus var. farinosus, 324
griseus var. purpureus, 140, 186, 187
griseus var. spiralis, 324
grisinus, 324
hachijoensis, 94, 149, 163, 185, 226, 227
hachijoensis var. fuscatus, 227
halstedizi, 93, 105, 160, 227, 252, 269
hawaiiensis, 145, 161, 227, 228
hepaticus, 217, 324
herbaricolor, 292
hiroshimensis, 101, 105, 109, 150, 151, 164,
228, 229
hirsutus, 93, 99, 130, 156, 229
hominis, 159, 229
humidus, 156, 229, 230
hygroscopicus, 68, 74, 98, 94, 101, 105, 107, 117,
124, 144, 160, 204, 230, 231, 259, 264
hygroscopicus var. angustmyceticus, 231
hygroscopicus var. decoyicus, 231
hygroscopicus var. odoratus, 231
intermedius, 123, 157, 231, 232, 286
tpomoeae, 78, 161, 174, 232
kanamyceticus, 133, 159, 232
kentuckensis, 163, 232, 233
kimbert, 156, 233
kitasatoensis, 164, 185, 233, 234
kitasawaensis, 161, 234
lanatus, 102, 162, 234
lavendulae, 29, 64, 74, 76, 77, 93, 94, 100, 102,
105, 108, 117, 146-148, 163, 196, 234, 235, 281,
287
lavendulae var. brasilicus, 235
lavendulae var. japonicus, 235
levoris, 325
leydenematis, 325
lieskei, 158, 232, 235
lilacinus, 325
limosus, 144, 158, 235, 236
lipmanit, 105, 160, 236, 268
litmocidini, 131
loidensis, 162, 236, 237
longisporoflavus, 160, 237, 254
longispororuber, 93, 252
longisporus, 325
lucensis, 161, 237
INDEX OF ORGANISMS 307
luridus, 133, 160, 237, 238
luteochromogenes, 325
luteolutescens, 325
luteoverticillatus, 101, 163, 238
lydicus, 158, 238, 239
macrosporeus, 99, 156, 239
maculatus, 157, 239
madurae, 27, 29, 159, 239, 240
marginatus, 161, 240
marinolimosus, 158, 240
marinus, 156, 240, 241
mashuensis, 163, 241
massasporeus, 292
matensis, 163, 241, 242
mediocidicus, 163, 242
mediterranet, 325
melanochr omogenes, 325
melanocyclus, 129, 160, 242, 243
melanogenes, 129, 162, 243
melanosporeus, 129
melanosporus, 325
michiganensis, 99, 102, 162, 243
microflavus, 105, 124, 129, 159, 169, 237, 243,
244, 270
mirabilis, 161, 222, 244
mitakaensis, 160, 244, 245
murinus, 102, 157, 245
naganishi, 94, 160, 245, 246
narbonensis, 158, 246, 252
natalensis, 325
netropsis, 100, 105, 151, 164, 246, 247
niger, 158, 247
nigrifaciens, 161, 247, 248
nigrificans, 144
nitrificans, 160, 248
nitrosporeus, 105, 158, 188, 248
niveoruber, 99, 129, 157, 248, 249
niveus, 123, 156, 249
noboritoensis, 162, 242, 249, 250
nodosus, 160, 250
noursel, 64, 99, 158, 250, 251
novaecaesareae, 131, 157, 251
odorifer, 163, 251
otdiosporus, 160, 251, 252
olivaceus, 76, 101, 102, 105, 131, 132, 159, 206,
Zi 223, 227,246, 252, 281
olivochromogenes, 94, 161, 222, 252, 253
olivoreticuli, 164, 253
olivoverticillatus, 101, 163, 253, 254
omiyaensis, 160, 254.
orchidaceus, 325
orientalis, 156, 254, 255, 265
ostreogriseus, 292
paraguayensis, 29, 159, 255
parvullus, 64, 101, 105, 123, 142, 157, 215, 255
parvus, 105, 124, 126, 127, 142, 157, 255, 25
paucisporogenes, 325
pelletiert, 27, 29, 159, 256
pentaticus, 164, 256, 257
phaeochromogenes, 64, 93, 99, 105, 162, 167,
Dad 24a 2005 2D0
phaeochromogenes var. chloromyceticus, 281
phaeofaciens, 228, 325
phaeopurpureus, 101, 162, 257, 258
phaeoviridis, 157, 258
phoenix, 325
pilosus, 99, 161, 258
platensis, 144, 156, 231, 258, 259
pleofaciens, 325
plicatus, 325
pluricolor, 131, 157, 259
pluricolorescens, 158, 259, 260, 325
poolensis, 162, 260
praecox, 159, 260
praefecundus, 162, 261
prastnoptlosus, 93, 99, 130, 156, 261
prasinus, 93, 99, 130, 156, 261
promycint, 3825
psammoticus, 292
pseudogriseolus, 159, 261, 262
puniceus, 141, 186
purpeofuscus, 94, 200
purpurascens, 64, 93, 99, 105, 106, 130, 161,
182, 194, 263
purpureochromogenes, 94, 129, 162, 166, 200,
234, 262, 263
pur pureofuscus, 162, 262
purpureus, 186
putrificus, 159, 263, 264
pyridomyceticus, 160, 264
racemochromogenus, 325
raffinosus, 325
rameus, 161, 264, 265
ramnait, 158, 265
ramulosus, 65, 101, 158, 265, 266
recifer, 325 n
rectus, 302, 303
resistomycificus, 161, 222, 266
reticuli, 64, 94, 100, 102, 105, 111, 117, 150, 151,
164, 210, 234, 266, 267
reticuli var. latumctdicus, 267
reticuloruber, 150
rumosus, 102, 124-127, 157, 173, 176, 204, 267
rimosus f. paramomycinus, 267
rochei, 159, 212, 267, 268
roseochromogenes, 93, 105, 108, 129, 161, 196,
204, 268
roseocitreus, 162, 269
roseodiastaticus, 160, 269
roseoflavus, 129, 159, 269, 27(
roseoverticillatus, 101, 164, 27(
roseus, 133, 159, 270, 271
INDEX OF ORGANISMS
ruber, 94, 102, 105, 117, 129, 159, 225, 252, 271
rubescens, 160, 271, 272
rubrireticult,
212
rutgersensis,
rulgersensis
99, 150, 164, 2
21, 229, 257, 270,
105, 160, 231, 250, 272, 273
var. castelarense,
sahachirot, 157, 273
sakatensis, 325
salmonicida,
105, 151, 325
sampsonit, 157, 273, 275
sayamaensis, 157, 273, 274
scabies, 71,
260, 274, 2
setonit. 158, 2
sindenensis,
78, 101, 105, 117,
75
275
geo
elie 159 1276
spheroides, 156, 2 276, 27
spiralis, 158, 242, 275, 277
sprroverticillatus, 101, 164,
subtropicus,
325
231, 325
144, 145, 161, 252,
7, 278
sulphureus, 105, 157, 278, 279
tanashiensis,
tendae, 100,
163, 279
tenuis, 161, 279
termitum, 322
thermodiastaticus, 302, 303, 320
thermofuscus, 301, 802, 303
thermophilus, 301, 303
thermoviolaceus, 302, 304
thermoviolaceus apingens, 304
thermoviolaceus pingens, 304
thermovulgar
7s, 302, 304
105, 162, 250, 279
thioluteus, 94, 105, 151, 163, 279, 280
toxicus, 325
toyocaensis,
tricolor, 130
tumuli, 156,
325
280
tyrosinaticus, 212
vendargus, 3
25
venezuelae, 66, 68, 93, 100, 105, 146, 147, 163,
280, 281
venezuelae var.
verne, 105, 157,
verticillatus,
verticillus, 3:
252, 281
roseosport, 281
150, 151, 163, 238, 281, 282
verticilloviridans, 150
25
vinaceous, 105
vinaceus, 105, 141,
vinaceus-drappus, 325
violaceochromogenes, 131
violaceoniger,
282
violaceoruber, 78, 93, 117, 130-132, 157, 184,
197, 198, 212, 251, 2
violaceus, 67
186, 260, 262
66, 100, 105, 130, 144, 157, 231,
59, 282-284.
, 157, 184, 212, 284, 285
violaceus-niger, 282
virgatus, 157, 285
virginiae, 105, 147, 148, 163, 196, 235, 285, 286
viridans, 129, 157, 286
viridifaciens, 325
viridis, 117, 180, 160, 184, 286, 287, 290
viridochromogenes, 67, 93,99, 101, 105, 117, 180,
149, 150, 162, 193, 215, 287, 288, 289
viridoflavus, 164, 215, 288-290
viridogenes, 66, 101, 161, 290
viridosporus, 325
vulgaris, 325
wedmorensis, 159, 290
willmoret, 156, 291
xanthochromogenes, 325
canthophaeus, 93, 157, 265, 291, 292
zaomyceticus, 325
Streptomycetaceae, vil, 2
Streptosporangium, Vil, 5, 81, 84, 310, 311, 313, 314
album, 314
amethystogenes, 314
roseum, 312, 313, 314
viridialbum, 314
vulgare, 314
Streptothrix, 10, 144, 315, 316
actinomyces, 14, 316
actinomycotica, 14, 316
alba, 6, 118, 119, 120, 122
albido, 316
albido-flava, 128
alpha, 316
aquatilis, 316
aurea, 317
beta, 317
bovis communis, 317
candida, 119, 122, 187, 317
caprae, 38, 317
chromogena, 118, 128, 144
cinereonigeraromaticus, 317
citrea, 123
coelicolor, 140, 197
cunicult, 57, 318
dassonvillei, 122, 123
eppingert, 35, 318
erythrea, 318
farcinica, 318
flava, 1
Foersterz, 118, 119, 318
foersteri, 123
forsteri, 318
freerz, 318
gedanensis, 119
gelatinosus, 319
graminearum, 123
hominis, 229, 319
humifica, 319
INDEX OF ORGANISMS 309
israeli, 14, 17, 319 violacea, 131
lathridizi, 119, 123, 319 zopfi, 322
leucea, 123, 319 Streptoverticillium, 5, 96, 111, 150
leucea saprophytica, 319 Thermoactinomyces, vit, 2, 5, 84, 301, 302, 306-309
melanotica, 320 glaucus, 3807
murts-rattt, 49 monosporus, 307, 309
necrophora, 320 thalpophilus, 307, 308
nigra, 49, 320 thermophilus, 307, 308
nigrescens, 320 viridis, 300, 307, 308, 309
oidioformis, 320 vulgaris, 307, 309
orangica, 321 Thermomonospora, vit, 5, 302, 304, 305, 306, 307
paulotrophus, 321 curvata, 304, 305
putoriz, 321 fusca, 804, 305
pyogenes, 123, 321 lineata, 304, 305, 306
rattt, 321 Thermophilic actinomycetes, 300-309
rubea, 321 Thermopolyspora, vii, 5, 302, 305, 306
rubra, 321 bispora, 306
sanninit, 322 polyspora, 306
spirilloides, 322 Waksmania, 2, 5, 84, 298, 299
taraxeri-cepapt, 322 rosea, 298, 299
tartart, 322 See also Microbispora
GENERAL INDEX
Abikoviromycin, 166, 272
Acetomyein, 265
Achromoviromyein, 167
Acidomyein, 320
Actinoidin, 324
Actinolysin, 324
Actinomyces
classification of, 12
description of, 12-19
genus of, 12-20
incompletely described forms of, 20, 316-323
species of, 13-20
Actinomycetin, 172
Actinomycin, 174, 192, 194, 209, 210, 211, 215,
234, 243, 245, 255, 324
Actinomycin-producing organisms, 141, 142
Actinomycosis, 78
Actinophage sensitivity of Streptomyces,
141, 142
Actithiazie acid, 196, 286
Aerial mycelium, 62-69
Aggregate-species, 9
Alazopeptin, 224, 324
Albomyein, 325
Allomyein, 825
Althiomyein, 173
Amicetin, 324, 325
Aminocidin, 324
Amphomyein, 190
Amphotericin, 250
Amylocyanin, 140
Anaerobie actinomycetes,
=p
75-76,
12-20
Angolamyein, 205
Anisomyein, 223
Antagonisms between strains, 147
Antibacterial properties, 147
Antibioties
1968, 324
C and D, 325
produced by
323-325
production of, as species characteristic, 75
incompletely deseribed species,
sensitivity of actinomycetes to, 74, 75
X, 324
Antibiotic-producing actinomycetes, 324-325
Antitrichomonal activity, 225, 262
Antitumor agent, 325
substances, 324
Arthrospores, 121
Asecosin, 190
Aspartoein, 324
Aureothricin, 192, 200, 28
Azaserine, 213
Bacteria, relationship to actinomycetes, v, 1
Bennett’s agar, 331, 332
Bergey’s Manual of Determinative Bacteriology, v
326
Biochemical properties of Streptomyces, 61
Biomyein, 325
Blasticidin, 221
Blastmyein, 182, 324
Blood agar, 334
B-mycin, 183
Borrelidin, 268
Bryamyein, 228
,
Cacaomycetin, 183
Caerulomyein, 184
Camphomyein, 325
Candicidin, 227
Candidin, 290
Candimycin, 324
Carbomyein, 227, 239, 279
Carbon nutrition medium, 332
Carbon sources for actinomycetes, 71-73, 143
Carcinomycin, 324
Carrot plug, 334
Caryomycin, 207
Carzinocidin, 234, 324
Carzinophilin, 273
Catenulin, 191
Celesticetin, 184
Cell wall, composition of, 1, 22
Celloeidin, 193, 324
Cellostatin, 324
Cellulose medium, 332
Chartreusin, 193
Chemical composition of actinomycetes,
Chitin, |
Chlamydospores, 298
Chloramphenicol, 254, 281
Chlortetracycline, 177, 274, 325
Cinerubin, 182, 249
Cinnamyein, 195
Cladomyein, 325
Classification systems of genus Streptomyces, 82-
114
Baldacei, 94-96
Belin ger et al.,
|
~J
98-101
360
GENERAL INDEX
Flaig and Kutzner, 93
Frommer, 101-102
Gause et al., 96-97
Hesseltine ef al., 92, 93
Jensen, 89
KrassilInikov, 89-91
Mayama, 102, 103
Nomi, 108-105
Pridham et al., 97, 98
Proposed new system for series, 110-114
Routien’s outline, 105
Shinobu, 101
Waksman, 87-89
Waksman and Curtis, 86, 87
Waksman and Henrici, 91
Yamaguchi and Saburi, 94
Cluster formation, 10, 66
Cohn, F., 0
Colimyein (probably neomycin), 324
Colony structure of streptomycetes, 69, 70
Color designations, 327, 328
Congocidin, 173
Conn’s medium, 329
Coremia formation, 68, 298
Croceomycin, 324
Crystallomycin, 325
Cyanomycin, 200
Cycloheximide, 140, 194, 226
Cycloserine, 325
Czapek’s agar, 328
Dextrin-casein digest agar, 332
Dorset’s medium, 334
Echinomycin, 208
Ecology of actinomycetes, 77, 78
Egg-albumin agar, 331
Ehrlichin, 235
Elaiomyein, 217
Elaiophylin, 325
Kmerson’s agar, 331
Endomycein, 172, 204
Erythromycin, 204, 205
Etamycin, 225
Etruscomycin, 237
Eurocidin, 169, 205
Fairy rings, 298
Fermicidin, 22:
Fervenulin, 207
Filipin, 208
Flavensomycin, 191
Flavofungin, 324
Flavomycin, 270
Fluorin, 324
Fradicin, 212
Fungichromin, 192
Fungicidin, 214
Ganmycin, 3824
Gelatin agar medium, 330
Gelatin media, 330
Genera of actinomycetes, 2, 82-84
Genetics of actinomycetes, 78
Geomyein, 291
Glucose agars
glucose-ammonium salt agar, 329
-asparagine agar, 328
-casein digest-yeast-beef agar, 331
-peptone agar, 329
-tyrosine agar, 329
-yeast-ammonium agar, 330
-yeast extract agar, 331
-yeast extract-beef-peptone agar, 331
-yeast-malt agar, 332
Glycerol agars
glycerol-ammonium salt agar, 329
-asparaginate agar, 328
-Camalate agar, 329
-glycine agar, 329
-peptone-beef agar, 330
-starch-glutamate agar, 331
-urea agar, 329
-yeast-malt agar, 640
Granaticin, 252
Grisamine, 222
Grisein, 220
Griseoflavin, 222
Griseolutein, 223
Griseomycin, 223
CGriseoviridin, 225
Grisin, 324
Groups of Streptomyces, 93
Hehomyein, 620
Helixin, 175
Hickey and Tresner’s agar, 332
Homomycin, 250
Hydrogen sulfide medium, 332
Hydroxystreptomycin F221
Hygromycein, 231, 250
Incompletely described species, 11, 215-226
Kanamycin, 232
Leucomycin, 234
Levorin, 325
Longisporin, 325
Luteomycin, 179, 279
Mangel scab, 280
Masumoto agar, 329
361
362
Matamycin, 180, 242
Media
composition of, 62, 328-334
standard, 80
Mediocidin, 242
Melanin
formation, 71, 333
-negative species, 111, 112
-positive species, 112
Mesophilic streptomycetes, 111, 112
Metabolism of Streptomyces griseus, 139, 140
Microbial species, concept of, 2-4
Micromonosporin, 296
Mikamyein, 245
Milk medium, 332
Miramyein, 244
Mitomycin, 185
Monilin, 325
Monomyein, 324
Morphology of streptomycetes, 5, 7, 61, 62-69, 116
Mutants of Streptomyces griseus, 140-143
Mycomycin, 324
Natural classification, 4
Necrotin, 325
Neomycin, 170, 212, 232, 266, 270
Nitrate reduction medium, 332
Nitrogen sources, 74
Nitrosporin, 248
Nocardia, 21-60, 323
acid-fast forms, 24, 27
biochemical properties of, 26-28
characterization of, 21-23
classification of, 28-34
colonies of, 24
description of, 27-29, 34-60
growth characteristics of, 24, 25
morphology of, 21-28
motility, 22
multiplication of, 21
serological properties, 29
Noecardorubin, 324
Noeardiosis, 21
Nomenclatural taxonomy, 4
Novobiocin, 216, 222, 249, 277
Nuelei, 4
Nucleoeidin, 187
Nutrient agar, 330
Nystatin, 251
Oatmeal agar, 332
Olivacein, 252
Oxytetracycline, 176, 223, 259, 267, 325
Pathogenic actinomycetes, 29
Pentamyein, 257
GENERAL INDEX
Peptone-beef extract agar, 330
-beef-salt agar, 330
Phaeofacin, 325
Phagomyein, 223
Phthiomyein, 325
Physiological classification, 6
Picromycein, 206, 246
Pigments, nature of, 70, 71
Pimaricin, 325
Pleomyein, 825
Plotho’s agar, 329
Pluramyein, 260, 325
Pneumocin, 325
Polyenes, 179, 198
Potato glucose agar, 331
-peptone-glycerol agar, 331
Potato
plug, 332
seab, 273, 275, 277
**Pox,:” 260
Proactinomycin, 216, 248
Progesterone, 176
Proteolytic activities of Streptomyces, 73, 74
Puromyein, 171
Pyridomycin, 264, 324
Racemomycin, 325
Raisnomyein, 2383
Ramnacin, 265
Reducing properties, 74
Resistomycin, 266
Rhodoeidin, 325
Rhodomyein, 194, 263
Rifomyein, 325
Rimocidin, 267
Ristocetin, 324
Roseocitrin, 269
Roseomycin, 268
Ruticin, 27:
Sabouraud’s agar, 329
Sarkomyein, 205
Sclerotia formation, 69, 70
Sections, 9, 97, 98
Rectus-flexibilis, 97
Retinaculum-apertum, 97
Spira, 97
Monoverticillus, 97
Monoverticillus-spira, 97
Biverticillus, 97
Biverticillus-spira, 97
Series concept, 9
Series of Streptomyces, 115-151
Serological reactions, 76, 77
Sistomycosin, 825
Soil-extract agar, 332
Speciation of streptomycetes, 6-10
Species
concept, I-11
descriptions, requirements for, LO, 11
groups, 7, 9, 66
Spiral types, 63-66
Spiramyein, 173
Spores of Streptomyces, 67-69
Starch agar, 330
-casein agar, 330
-peptone-beef agar, 331
Steroids, 266
Streptin, 272
Streptogramin, 324
Streptolin, 324
Streptomyces
characterization of, 61-81, 153-155
classification of, 85-105, 155-164
description of, 10, 80, 81, 84, 165-292
generic name, 2
morphological groups, 104
series of, 115-151
species concept of, 2-4
species of, 84, 152-164
Streptomycetes
actinophage sensitivity, 75, 76
antibiotic sensitivity, 74, 75
biochemical properties, 70-74
earbon utilization, 71-73
chemical composition, 77
colony structure, 69, 70
cultural properties, 70-74
ecology, 77, 78
genera of, 82-84
genetics, 78
growth response, 333
nitrogen utilization, 74
pigment formation, 70, 71
proteolytic properties, 73, 74
reducing properties, 74
serological reactions, 76, 77
standard media, 80
type cultures, 78-80
Streptomycin
-dependent strains, 136
GENERAL INDEX
-producing strains, 136, 141
production, 142, 143, 181, 226, 241, 252, 2
Streptonivicin, 249
Streptothricin, 169, 235, 244, 324
Streptovaricin, 276
Substrate mycelium, 62
Sucrose-nitrate agar, 328
Synthetic media, 7
Systematic position of actinomycetes, 1
Taitomycin, 168
Taxonomy of actinomycetes, 4
Tennecetin, 324
Tetracycline, 325
>)
»-
Thermophilic actinomycetes, 112, 300-309
Thiozolidone, 196
Tomato paste-oatmeal agar, 332
Toyocamyein, 325
Trichomycin, 227
Trichonin, 272
Tryptone-yeast agar, 330
Tuft formation, 10
Tumor-inhibiting substance, 243
Type cultures, 10, 61, 78-80
Tyrosinase reaction, 9, 71
Tyrosine-casein-nitrate agar, 329, 330
Tundramyein, 324
Ushinsky’s medium, 328
. >?
Valinomycin, 213
Vancomycin, 254
Variations, cultural, 72
Varieties of Streptomyces griseus, 140-148
Vengicide, 325
65
363
Verticil-forming species of Streptomyces, 64, 65
Viomyein, 140, 186, 187, 253
Viridogrisein, 225
Virocidin, 210
Vitamin Bie , 824
Xanthicin, 325
Xanthomycin, 262, 273
Zaomycin, 325
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